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antibody against fzd7  (R&D Systems)


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    R&D Systems antibody against fzd7
    Schematic representation of the <t>FZD7</t> protein structure and its localization within the cell membrane. Four peptides were selected for antibody generation; clones 288.1, 288.2, and 288.5 derived from peptide 4 were subsequently chosen for detailed analysis. Figure created with BioRender.com.
    Antibody Against Fzd7, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/antibody+against+fzd7/Human%2FMouse+Frizzled-7+APC-conjugated+Antibody/pmc12504202-228-18-21
    Average 93 stars, based on 4 article reviews
    antibody against fzd7 - by Bioz Stars, 2026-10
    93/100 stars

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    1) Product Images from "A novel Frizzled 7 antibody disrupts the Wnt pathway and inhibits Wilms tumor growth"

    Article Title: A novel Frizzled 7 antibody disrupts the Wnt pathway and inhibits Wilms tumor growth

    Journal: Frontiers in Bioengineering and Biotechnology

    doi: 10.3389/fbioe.2025.1641137

    Schematic representation of the FZD7 protein structure and its localization within the cell membrane. Four peptides were selected for antibody generation; clones 288.1, 288.2, and 288.5 derived from peptide 4 were subsequently chosen for detailed analysis. Figure created with BioRender.com.
    Figure Legend Snippet: Schematic representation of the FZD7 protein structure and its localization within the cell membrane. Four peptides were selected for antibody generation; clones 288.1, 288.2, and 288.5 derived from peptide 4 were subsequently chosen for detailed analysis. Figure created with BioRender.com.

    Techniques Used: Membrane, Clone Assay, Derivative Assay

    Binding of FZD7 antibody to Wilms tumor cells, expression analysis of different WT donors and effect on cell proliferation. (A) Percentage of Frizzle 7 (FZD7)-expressing cells in culture. A representative flow cytometry analysis is shown for primary Wilms tumor (WT)-derived cells (upper) and WT-xenograft derived cells (lower). (B) A summarizing bar graph of FZD7 expression in WT cells from different donors used for in vitro experiments. (C) Effect of secreted antibodies on WT cells: Relative percentage of viable cells following 48 h of treatment with the different clones. The spent medium served as a control. The numbers indicate the hybridoma clone from which the spent medium was derived; *p < 0.05, **p < 0.01. The results are shown as the mean ± SEM of at least three different experiments for each antibody.
    Figure Legend Snippet: Binding of FZD7 antibody to Wilms tumor cells, expression analysis of different WT donors and effect on cell proliferation. (A) Percentage of Frizzle 7 (FZD7)-expressing cells in culture. A representative flow cytometry analysis is shown for primary Wilms tumor (WT)-derived cells (upper) and WT-xenograft derived cells (lower). (B) A summarizing bar graph of FZD7 expression in WT cells from different donors used for in vitro experiments. (C) Effect of secreted antibodies on WT cells: Relative percentage of viable cells following 48 h of treatment with the different clones. The spent medium served as a control. The numbers indicate the hybridoma clone from which the spent medium was derived; *p < 0.05, **p < 0.01. The results are shown as the mean ± SEM of at least three different experiments for each antibody.

    Techniques Used: Binding Assay, Wilms Tumor Assay, Expressing, Flow Cytometry, Derivative Assay, In Vitro, Clone Assay, Control

    Binding characterization of FZD7 -288.1 in Immune Precipitation assays, Western blot analysis, FACS and Immunohistochemistry. (A) αFZD7-288.1 but not 288.5 immunoprecipitated the Frizzled 7 (FZD7) protein. A protein immunoprecipitation (IP) assay was used to validate the antibody-specific interaction with the FZD7 receptor. SK-MEL28 total cell lysates (FZD7 Input) and FZD7-bound fractions (FZD7 IP) were assayed via Western blot analysis. For detection, a commercial anti-FZD7 antibody (Millipore) was used. Nonspecific antibody and mouse IgG served as negative controls, and β-actin was used as a loading control. A representative blot of three separate experiments is shown. (B) Specific binding to the FZD7 receptor was examined in HEK293T cells overexpressing full-length FZD7. The cell extracts were probed with different concentrations of αFZD7-288.1. Specific binding of the Ab to FZD7-expressing cells was observed. (C) Probing of cell lysates from sh-FZD7-Wilms tumor (WT) cells with αFZD7-288.1 revealed the downregulation of FZD7, similar to the effect of a commercial αFZD7 antibody, demonstrating its specificity. β-Actin was used as a loading control. (D) Representative flow cytometry analysis of FZD7 expression in cultured WT cells stained with both αFZD7-288.1 and a rat anti-FZD7 APC-conjugated antibody. Cells stained positive for αFZD7-288.1 were mostly observed as a subpopulation within the cells detected by RαFZD7-APC. The data are presented in a dot plot graph showing FZD7 staining in red and isotype control staining (negative control) in gray.
    Figure Legend Snippet: Binding characterization of FZD7 -288.1 in Immune Precipitation assays, Western blot analysis, FACS and Immunohistochemistry. (A) αFZD7-288.1 but not 288.5 immunoprecipitated the Frizzled 7 (FZD7) protein. A protein immunoprecipitation (IP) assay was used to validate the antibody-specific interaction with the FZD7 receptor. SK-MEL28 total cell lysates (FZD7 Input) and FZD7-bound fractions (FZD7 IP) were assayed via Western blot analysis. For detection, a commercial anti-FZD7 antibody (Millipore) was used. Nonspecific antibody and mouse IgG served as negative controls, and β-actin was used as a loading control. A representative blot of three separate experiments is shown. (B) Specific binding to the FZD7 receptor was examined in HEK293T cells overexpressing full-length FZD7. The cell extracts were probed with different concentrations of αFZD7-288.1. Specific binding of the Ab to FZD7-expressing cells was observed. (C) Probing of cell lysates from sh-FZD7-Wilms tumor (WT) cells with αFZD7-288.1 revealed the downregulation of FZD7, similar to the effect of a commercial αFZD7 antibody, demonstrating its specificity. β-Actin was used as a loading control. (D) Representative flow cytometry analysis of FZD7 expression in cultured WT cells stained with both αFZD7-288.1 and a rat anti-FZD7 APC-conjugated antibody. Cells stained positive for αFZD7-288.1 were mostly observed as a subpopulation within the cells detected by RαFZD7-APC. The data are presented in a dot plot graph showing FZD7 staining in red and isotype control staining (negative control) in gray.

    Techniques Used: Binding Assay, Western Blot, Immunohistochemistry, Immunoprecipitation, Control, Expressing, Wilms Tumor Assay, Flow Cytometry, Cell Culture, Staining, Negative Control

    The αFZD7-288.1 antibody inhibits β-catenin translocating Wnt signaling in Wilms tumor cells. (A) Western blot (WB) analysis demonstrating the inhibition of active β-catenin and decreased levels of Fizzled 7 (FZD7). (B) following treatment with 5 μg/mL αFZD7-288.1 for 48 h in Wilms tumor (WT) cells. A nonspecific Ab was used as a negative control; β-actin was used as a loading control. (C) β-catenin immunofluorescence staining of untreated WT and αFZD7-288.1-treated cells: Double-staining with DAPI and cytokeratin (first panel) and with cytokeratin and β-catenin (second panel). β-catenin staining revealed strong expression and a greater percentage of cells with nuclear localization of β-catenin in the control untreated cells (scale bar represents 50 µM) X70 magnification of representative cells (indicated by arrows) from each group (third panel) and double-staining with DAPI and β-catenin (forth panel). (D–F) Real-time PCR gene expression analysis of WT cells: The expression of Wnt transducers [ (D) ; FZD7, β-catenin], Wnt pathway target genes [ (E) ; AXIN2, CCND1, MYC-C], and Wnt pathway inhibitors [ (F) ; DKK1, sFRP1] was also measured. The results showed reduced mRNA expression of Wnt pathway-related genes. The data are shown as the mean ± S.E.M. of 5 separate experiments; *p < 0.05; **p < 0.01.
    Figure Legend Snippet: The αFZD7-288.1 antibody inhibits β-catenin translocating Wnt signaling in Wilms tumor cells. (A) Western blot (WB) analysis demonstrating the inhibition of active β-catenin and decreased levels of Fizzled 7 (FZD7). (B) following treatment with 5 μg/mL αFZD7-288.1 for 48 h in Wilms tumor (WT) cells. A nonspecific Ab was used as a negative control; β-actin was used as a loading control. (C) β-catenin immunofluorescence staining of untreated WT and αFZD7-288.1-treated cells: Double-staining with DAPI and cytokeratin (first panel) and with cytokeratin and β-catenin (second panel). β-catenin staining revealed strong expression and a greater percentage of cells with nuclear localization of β-catenin in the control untreated cells (scale bar represents 50 µM) X70 magnification of representative cells (indicated by arrows) from each group (third panel) and double-staining with DAPI and β-catenin (forth panel). (D–F) Real-time PCR gene expression analysis of WT cells: The expression of Wnt transducers [ (D) ; FZD7, β-catenin], Wnt pathway target genes [ (E) ; AXIN2, CCND1, MYC-C], and Wnt pathway inhibitors [ (F) ; DKK1, sFRP1] was also measured. The results showed reduced mRNA expression of Wnt pathway-related genes. The data are shown as the mean ± S.E.M. of 5 separate experiments; *p < 0.05; **p < 0.01.

    Techniques Used: Wilms Tumor Assay, Western Blot, Inhibition, Negative Control, Control, Immunofluorescence, Staining, Double Staining, Expressing, Real-time Polymerase Chain Reaction, Gene Expression

    The αFZD7-288.1 antibody reduces proliferation in Wilms tumors cells and induces cell death. (A) Inhibition of the Wnt pathway by αFZD7-288.1 treatment induced changes in cell kinetics, as demonstrated by the longer doubling time of antibody treated Wilms tumor (WT) cells compared to control treated WT cells and (B) the significantly lower growth rate (defined as the number of cell divisions per day), *p < 0.05; n = 4. Reduced proliferation in cells treated with αFZD7-288.1 was also observed by MTS assay in (C) and (D) Quantification of Ki67 staining was performed by counting the number of Ki67-positive nuclei in five high-power fields (HPF) per condition, using representative regions from each sample. Reduced nuclear Ki67 expression (red, lower panels) compared to that in control untreated cells (magnification ×40 scale bars = 100 μm; magnification ×20 scale bars = 50 µm). (E) Induction of cell death in WT cells is shown in a representative analysis for annexin V staining, which demonstrated a marked increase in the percentage of apoptotic and preapoptotic cells. (F) Percentage of dead cells calculated using trypan blue staining of WT cells indicating significantly greater cell death in treated WT cells than in control cells; *p < 0.05; n = 4.
    Figure Legend Snippet: The αFZD7-288.1 antibody reduces proliferation in Wilms tumors cells and induces cell death. (A) Inhibition of the Wnt pathway by αFZD7-288.1 treatment induced changes in cell kinetics, as demonstrated by the longer doubling time of antibody treated Wilms tumor (WT) cells compared to control treated WT cells and (B) the significantly lower growth rate (defined as the number of cell divisions per day), *p < 0.05; n = 4. Reduced proliferation in cells treated with αFZD7-288.1 was also observed by MTS assay in (C) and (D) Quantification of Ki67 staining was performed by counting the number of Ki67-positive nuclei in five high-power fields (HPF) per condition, using representative regions from each sample. Reduced nuclear Ki67 expression (red, lower panels) compared to that in control untreated cells (magnification ×40 scale bars = 100 μm; magnification ×20 scale bars = 50 µm). (E) Induction of cell death in WT cells is shown in a representative analysis for annexin V staining, which demonstrated a marked increase in the percentage of apoptotic and preapoptotic cells. (F) Percentage of dead cells calculated using trypan blue staining of WT cells indicating significantly greater cell death in treated WT cells than in control cells; *p < 0.05; n = 4.

    Techniques Used: Inhibition, Wilms Tumor Assay, Control, MTS Assay, Staining, Expressing

    The migration and sphere formation of Wilms tumor cells is reduced after treatment with αFZD7-288.1 antibody. (A) Scratch assay demonstrating the effects of αFZD7-288.1 on the ability of cells to migrate through a scrape in the cell monolayer. Wilms tumor (WT)-treated cells demonstrated a lower migration capacity than control cells, as shown in the representative images. Calculation of the area of migrating cells (on the right) showed that the cells had a significantly reduced migration capacity following treatment; *p < 0.05; n = 3. (B) A sphere formation assay was performed. The number and size of the spheres formed by the treated cells were significantly lower than those formed by the untreated cells. The data are presented as the means ± SEMs of at least three experiments performed in triplicate (*p < 0.05) On the left, representative phase-contrast images of spheres formed by treated and untreated WT cells showing markedly fewer condensed spheres in the treated cells (scale bars = 100 μm, magnification ×10).
    Figure Legend Snippet: The migration and sphere formation of Wilms tumor cells is reduced after treatment with αFZD7-288.1 antibody. (A) Scratch assay demonstrating the effects of αFZD7-288.1 on the ability of cells to migrate through a scrape in the cell monolayer. Wilms tumor (WT)-treated cells demonstrated a lower migration capacity than control cells, as shown in the representative images. Calculation of the area of migrating cells (on the right) showed that the cells had a significantly reduced migration capacity following treatment; *p < 0.05; n = 3. (B) A sphere formation assay was performed. The number and size of the spheres formed by the treated cells were significantly lower than those formed by the untreated cells. The data are presented as the means ± SEMs of at least three experiments performed in triplicate (*p < 0.05) On the left, representative phase-contrast images of spheres formed by treated and untreated WT cells showing markedly fewer condensed spheres in the treated cells (scale bars = 100 μm, magnification ×10).

    Techniques Used: Migration, Wilms Tumor Assay, Wound Healing Assay, Control, Tube Formation Assay

    αFZD7-288.1 strongly co-localizes with NCAM1 in WT, and treatment with αFZD7-288.1 reduces WT CSC markers. (A) Immunofluorescence of staining of cultured WT cells using αFZD7-288.1 and NCAM1 and co-staining (αFZD7-red, αNCAM1 green and DAPI blue) microscopy revealed overlapping signals as a merged color (yellow), indicating membrane spatial proximity. Slides were imaged using Olympus IX83 fluorescent microscope and Olympus DP80 camera. Image processing was done using Olympus OlyVia software. (B) The expression of the Cancer stem cell marker genes NCAM1, CITED, SIX2 and ALDH1A1 decreased in the antibody αFZD7-288.1 treated WT derived cells (n = 3); *p < 0.05; **p < 0.01.
    Figure Legend Snippet: αFZD7-288.1 strongly co-localizes with NCAM1 in WT, and treatment with αFZD7-288.1 reduces WT CSC markers. (A) Immunofluorescence of staining of cultured WT cells using αFZD7-288.1 and NCAM1 and co-staining (αFZD7-red, αNCAM1 green and DAPI blue) microscopy revealed overlapping signals as a merged color (yellow), indicating membrane spatial proximity. Slides were imaged using Olympus IX83 fluorescent microscope and Olympus DP80 camera. Image processing was done using Olympus OlyVia software. (B) The expression of the Cancer stem cell marker genes NCAM1, CITED, SIX2 and ALDH1A1 decreased in the antibody αFZD7-288.1 treated WT derived cells (n = 3); *p < 0.05; **p < 0.01.

    Techniques Used: Immunofluorescence, Staining, Cell Culture, Microscopy, Membrane, Software, Expressing, Marker, Derivative Assay

    The antitumor activity of αFZD7-288.1 as demonstrated in changes in tumor volumes. (A) Flow cytometry histogram evaluating Frizzled 7 (FZD7) expression in Wilms tumor (WT) tissue for use in vivo . (B) Workflow diagram illustrating the in vivo experimental design. (C) Mouse weights during treatment. Paclitaxel (PTX)-treated mice suffered from toxic side effects and significant weight loss following treatment; **p < 0.01; ***p < 0.005. (D) Antitumor activity: Mice were treated intravenously with normal saline, αFZD7-288.1 (10 mg/kg) or paclitaxel (15 mg/kg). Tumor volume, presented in mm 3 , was assessed every 2–3 days using an external electronic caliper and calculated by the modified ellipsoid formula V = 0.52 × (length × width 2 ). Like PTX-treated tumors, tumors treated with αFZD7-288.1 exhibited growth inhibition and even tumor shrinkage, while saline-treated tumors showed rapid tumor growth; *p < 0.05; **p < 0.01. (E) Representative images on day 10 of the experiment showing mice with large saline-treated tumors (right) and unnoticeable αFZD7-288.1-treated tumors (left). (F) Waterfall graph showing the changes in the individual tumor volumes with respect to the initial tumor size. (G) FZD7 expression in tumors treated with αFZD7-288.1 was significantly altered by 0.445-fold; *p < 0.05; n = 3 per group. (H) The expression of the canonical Wnt pathway genes FZD7, β-CATENIN, AXIN2, CCND1, C-MYC, DKK1, and sFRP1 decreased in the treated tumors (n = 4); *p < 0.05; **p < 0.01; ***p < 0.005.
    Figure Legend Snippet: The antitumor activity of αFZD7-288.1 as demonstrated in changes in tumor volumes. (A) Flow cytometry histogram evaluating Frizzled 7 (FZD7) expression in Wilms tumor (WT) tissue for use in vivo . (B) Workflow diagram illustrating the in vivo experimental design. (C) Mouse weights during treatment. Paclitaxel (PTX)-treated mice suffered from toxic side effects and significant weight loss following treatment; **p < 0.01; ***p < 0.005. (D) Antitumor activity: Mice were treated intravenously with normal saline, αFZD7-288.1 (10 mg/kg) or paclitaxel (15 mg/kg). Tumor volume, presented in mm 3 , was assessed every 2–3 days using an external electronic caliper and calculated by the modified ellipsoid formula V = 0.52 × (length × width 2 ). Like PTX-treated tumors, tumors treated with αFZD7-288.1 exhibited growth inhibition and even tumor shrinkage, while saline-treated tumors showed rapid tumor growth; *p < 0.05; **p < 0.01. (E) Representative images on day 10 of the experiment showing mice with large saline-treated tumors (right) and unnoticeable αFZD7-288.1-treated tumors (left). (F) Waterfall graph showing the changes in the individual tumor volumes with respect to the initial tumor size. (G) FZD7 expression in tumors treated with αFZD7-288.1 was significantly altered by 0.445-fold; *p < 0.05; n = 3 per group. (H) The expression of the canonical Wnt pathway genes FZD7, β-CATENIN, AXIN2, CCND1, C-MYC, DKK1, and sFRP1 decreased in the treated tumors (n = 4); *p < 0.05; **p < 0.01; ***p < 0.005.

    Techniques Used: Activity Assay, Flow Cytometry, Expressing, Wilms Tumor Assay, In Vivo, Saline, Modification, Inhibition

    The antitumor activity of αFZD7-288.1 as demonstrated by proliferative markers in Immunohistochemical staining. (A) Immunohistochemical staining for Ki67 and Caspase3 in representative tumors from the different experimental groups; saline, paclitaxel (PTX) and αFZD7-288.1 (type A, representing reduced-volume tumors; and type B, representing tumors with growth inhibition compared to saline-treated tumors). The scale bars represent 50 μm at a magnification of ×20. Compared with those of saline- or PTX-treated tumors, αFZD7-288.1-treated tumors exhibited reduced proliferation. An increase in apoptosis was observed in αFZD7-288.1- and PTX-treated tumors (B) .
    Figure Legend Snippet: The antitumor activity of αFZD7-288.1 as demonstrated by proliferative markers in Immunohistochemical staining. (A) Immunohistochemical staining for Ki67 and Caspase3 in representative tumors from the different experimental groups; saline, paclitaxel (PTX) and αFZD7-288.1 (type A, representing reduced-volume tumors; and type B, representing tumors with growth inhibition compared to saline-treated tumors). The scale bars represent 50 μm at a magnification of ×20. Compared with those of saline- or PTX-treated tumors, αFZD7-288.1-treated tumors exhibited reduced proliferation. An increase in apoptosis was observed in αFZD7-288.1- and PTX-treated tumors (B) .

    Techniques Used: Activity Assay, Immunohistochemical staining, Staining, Saline, Inhibition

    Related Articles

    Staining:

    Article Title: A novel Frizzled 7 antibody disrupts the Wnt pathway and inhibits Wilms tumor growth
    Article Snippet: The cells were then blocked with a mixture of FcR blocking reagent (Miltenyi Biotec) and human serum (1:1). .. The cells were stained with an isotype control (R and D Systems Cat# IC006A, RRID:AB_357254) or a primary antibody against FZD7 (R and D Systems Cat# FAB1981A, RRID:AB_2232278). .. Cell viability was tested using 7AAD viability staining solution (eBioscience).

    Control:

    Article Title: A novel Frizzled 7 antibody disrupts the Wnt pathway and inhibits Wilms tumor growth
    Article Snippet: The cells were then blocked with a mixture of FcR blocking reagent (Miltenyi Biotec) and human serum (1:1). .. The cells were stained with an isotype control (R and D Systems Cat# IC006A, RRID:AB_357254) or a primary antibody against FZD7 (R and D Systems Cat# FAB1981A, RRID:AB_2232278). .. Cell viability was tested using 7AAD viability staining solution (eBioscience).



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    Schematic representation of the <t>FZD7</t> protein structure and its localization within the cell membrane. Four peptides were selected for antibody generation; clones 288.1, 288.2, and 288.5 derived from peptide 4 were subsequently chosen for detailed analysis. Figure created with BioRender.com.
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    Image Search Results


    Schematic representation of the FZD7 protein structure and its localization within the cell membrane. Four peptides were selected for antibody generation; clones 288.1, 288.2, and 288.5 derived from peptide 4 were subsequently chosen for detailed analysis. Figure created with BioRender.com.

    Journal: Frontiers in Bioengineering and Biotechnology

    Article Title: A novel Frizzled 7 antibody disrupts the Wnt pathway and inhibits Wilms tumor growth

    doi: 10.3389/fbioe.2025.1641137

    Figure Lengend Snippet: Schematic representation of the FZD7 protein structure and its localization within the cell membrane. Four peptides were selected for antibody generation; clones 288.1, 288.2, and 288.5 derived from peptide 4 were subsequently chosen for detailed analysis. Figure created with BioRender.com.

    Article Snippet: The cells were stained with an isotype control (R and D Systems Cat# IC006A, RRID:AB_357254) or a primary antibody against FZD7 (R and D Systems Cat# FAB1981A, RRID:AB_2232278).

    Techniques: Membrane, Clone Assay, Derivative Assay

    Binding of FZD7 antibody to Wilms tumor cells, expression analysis of different WT donors and effect on cell proliferation. (A) Percentage of Frizzle 7 (FZD7)-expressing cells in culture. A representative flow cytometry analysis is shown for primary Wilms tumor (WT)-derived cells (upper) and WT-xenograft derived cells (lower). (B) A summarizing bar graph of FZD7 expression in WT cells from different donors used for in vitro experiments. (C) Effect of secreted antibodies on WT cells: Relative percentage of viable cells following 48 h of treatment with the different clones. The spent medium served as a control. The numbers indicate the hybridoma clone from which the spent medium was derived; *p < 0.05, **p < 0.01. The results are shown as the mean ± SEM of at least three different experiments for each antibody.

    Journal: Frontiers in Bioengineering and Biotechnology

    Article Title: A novel Frizzled 7 antibody disrupts the Wnt pathway and inhibits Wilms tumor growth

    doi: 10.3389/fbioe.2025.1641137

    Figure Lengend Snippet: Binding of FZD7 antibody to Wilms tumor cells, expression analysis of different WT donors and effect on cell proliferation. (A) Percentage of Frizzle 7 (FZD7)-expressing cells in culture. A representative flow cytometry analysis is shown for primary Wilms tumor (WT)-derived cells (upper) and WT-xenograft derived cells (lower). (B) A summarizing bar graph of FZD7 expression in WT cells from different donors used for in vitro experiments. (C) Effect of secreted antibodies on WT cells: Relative percentage of viable cells following 48 h of treatment with the different clones. The spent medium served as a control. The numbers indicate the hybridoma clone from which the spent medium was derived; *p < 0.05, **p < 0.01. The results are shown as the mean ± SEM of at least three different experiments for each antibody.

    Article Snippet: The cells were stained with an isotype control (R and D Systems Cat# IC006A, RRID:AB_357254) or a primary antibody against FZD7 (R and D Systems Cat# FAB1981A, RRID:AB_2232278).

    Techniques: Binding Assay, Wilms Tumor Assay, Expressing, Flow Cytometry, Derivative Assay, In Vitro, Clone Assay, Control

    Binding characterization of FZD7 -288.1 in Immune Precipitation assays, Western blot analysis, FACS and Immunohistochemistry. (A) αFZD7-288.1 but not 288.5 immunoprecipitated the Frizzled 7 (FZD7) protein. A protein immunoprecipitation (IP) assay was used to validate the antibody-specific interaction with the FZD7 receptor. SK-MEL28 total cell lysates (FZD7 Input) and FZD7-bound fractions (FZD7 IP) were assayed via Western blot analysis. For detection, a commercial anti-FZD7 antibody (Millipore) was used. Nonspecific antibody and mouse IgG served as negative controls, and β-actin was used as a loading control. A representative blot of three separate experiments is shown. (B) Specific binding to the FZD7 receptor was examined in HEK293T cells overexpressing full-length FZD7. The cell extracts were probed with different concentrations of αFZD7-288.1. Specific binding of the Ab to FZD7-expressing cells was observed. (C) Probing of cell lysates from sh-FZD7-Wilms tumor (WT) cells with αFZD7-288.1 revealed the downregulation of FZD7, similar to the effect of a commercial αFZD7 antibody, demonstrating its specificity. β-Actin was used as a loading control. (D) Representative flow cytometry analysis of FZD7 expression in cultured WT cells stained with both αFZD7-288.1 and a rat anti-FZD7 APC-conjugated antibody. Cells stained positive for αFZD7-288.1 were mostly observed as a subpopulation within the cells detected by RαFZD7-APC. The data are presented in a dot plot graph showing FZD7 staining in red and isotype control staining (negative control) in gray.

    Journal: Frontiers in Bioengineering and Biotechnology

    Article Title: A novel Frizzled 7 antibody disrupts the Wnt pathway and inhibits Wilms tumor growth

    doi: 10.3389/fbioe.2025.1641137

    Figure Lengend Snippet: Binding characterization of FZD7 -288.1 in Immune Precipitation assays, Western blot analysis, FACS and Immunohistochemistry. (A) αFZD7-288.1 but not 288.5 immunoprecipitated the Frizzled 7 (FZD7) protein. A protein immunoprecipitation (IP) assay was used to validate the antibody-specific interaction with the FZD7 receptor. SK-MEL28 total cell lysates (FZD7 Input) and FZD7-bound fractions (FZD7 IP) were assayed via Western blot analysis. For detection, a commercial anti-FZD7 antibody (Millipore) was used. Nonspecific antibody and mouse IgG served as negative controls, and β-actin was used as a loading control. A representative blot of three separate experiments is shown. (B) Specific binding to the FZD7 receptor was examined in HEK293T cells overexpressing full-length FZD7. The cell extracts were probed with different concentrations of αFZD7-288.1. Specific binding of the Ab to FZD7-expressing cells was observed. (C) Probing of cell lysates from sh-FZD7-Wilms tumor (WT) cells with αFZD7-288.1 revealed the downregulation of FZD7, similar to the effect of a commercial αFZD7 antibody, demonstrating its specificity. β-Actin was used as a loading control. (D) Representative flow cytometry analysis of FZD7 expression in cultured WT cells stained with both αFZD7-288.1 and a rat anti-FZD7 APC-conjugated antibody. Cells stained positive for αFZD7-288.1 were mostly observed as a subpopulation within the cells detected by RαFZD7-APC. The data are presented in a dot plot graph showing FZD7 staining in red and isotype control staining (negative control) in gray.

    Article Snippet: The cells were stained with an isotype control (R and D Systems Cat# IC006A, RRID:AB_357254) or a primary antibody against FZD7 (R and D Systems Cat# FAB1981A, RRID:AB_2232278).

    Techniques: Binding Assay, Western Blot, Immunohistochemistry, Immunoprecipitation, Control, Expressing, Wilms Tumor Assay, Flow Cytometry, Cell Culture, Staining, Negative Control

    The αFZD7-288.1 antibody inhibits β-catenin translocating Wnt signaling in Wilms tumor cells. (A) Western blot (WB) analysis demonstrating the inhibition of active β-catenin and decreased levels of Fizzled 7 (FZD7). (B) following treatment with 5 μg/mL αFZD7-288.1 for 48 h in Wilms tumor (WT) cells. A nonspecific Ab was used as a negative control; β-actin was used as a loading control. (C) β-catenin immunofluorescence staining of untreated WT and αFZD7-288.1-treated cells: Double-staining with DAPI and cytokeratin (first panel) and with cytokeratin and β-catenin (second panel). β-catenin staining revealed strong expression and a greater percentage of cells with nuclear localization of β-catenin in the control untreated cells (scale bar represents 50 µM) X70 magnification of representative cells (indicated by arrows) from each group (third panel) and double-staining with DAPI and β-catenin (forth panel). (D–F) Real-time PCR gene expression analysis of WT cells: The expression of Wnt transducers [ (D) ; FZD7, β-catenin], Wnt pathway target genes [ (E) ; AXIN2, CCND1, MYC-C], and Wnt pathway inhibitors [ (F) ; DKK1, sFRP1] was also measured. The results showed reduced mRNA expression of Wnt pathway-related genes. The data are shown as the mean ± S.E.M. of 5 separate experiments; *p < 0.05; **p < 0.01.

    Journal: Frontiers in Bioengineering and Biotechnology

    Article Title: A novel Frizzled 7 antibody disrupts the Wnt pathway and inhibits Wilms tumor growth

    doi: 10.3389/fbioe.2025.1641137

    Figure Lengend Snippet: The αFZD7-288.1 antibody inhibits β-catenin translocating Wnt signaling in Wilms tumor cells. (A) Western blot (WB) analysis demonstrating the inhibition of active β-catenin and decreased levels of Fizzled 7 (FZD7). (B) following treatment with 5 μg/mL αFZD7-288.1 for 48 h in Wilms tumor (WT) cells. A nonspecific Ab was used as a negative control; β-actin was used as a loading control. (C) β-catenin immunofluorescence staining of untreated WT and αFZD7-288.1-treated cells: Double-staining with DAPI and cytokeratin (first panel) and with cytokeratin and β-catenin (second panel). β-catenin staining revealed strong expression and a greater percentage of cells with nuclear localization of β-catenin in the control untreated cells (scale bar represents 50 µM) X70 magnification of representative cells (indicated by arrows) from each group (third panel) and double-staining with DAPI and β-catenin (forth panel). (D–F) Real-time PCR gene expression analysis of WT cells: The expression of Wnt transducers [ (D) ; FZD7, β-catenin], Wnt pathway target genes [ (E) ; AXIN2, CCND1, MYC-C], and Wnt pathway inhibitors [ (F) ; DKK1, sFRP1] was also measured. The results showed reduced mRNA expression of Wnt pathway-related genes. The data are shown as the mean ± S.E.M. of 5 separate experiments; *p < 0.05; **p < 0.01.

    Article Snippet: The cells were stained with an isotype control (R and D Systems Cat# IC006A, RRID:AB_357254) or a primary antibody against FZD7 (R and D Systems Cat# FAB1981A, RRID:AB_2232278).

    Techniques: Wilms Tumor Assay, Western Blot, Inhibition, Negative Control, Control, Immunofluorescence, Staining, Double Staining, Expressing, Real-time Polymerase Chain Reaction, Gene Expression

    The αFZD7-288.1 antibody reduces proliferation in Wilms tumors cells and induces cell death. (A) Inhibition of the Wnt pathway by αFZD7-288.1 treatment induced changes in cell kinetics, as demonstrated by the longer doubling time of antibody treated Wilms tumor (WT) cells compared to control treated WT cells and (B) the significantly lower growth rate (defined as the number of cell divisions per day), *p < 0.05; n = 4. Reduced proliferation in cells treated with αFZD7-288.1 was also observed by MTS assay in (C) and (D) Quantification of Ki67 staining was performed by counting the number of Ki67-positive nuclei in five high-power fields (HPF) per condition, using representative regions from each sample. Reduced nuclear Ki67 expression (red, lower panels) compared to that in control untreated cells (magnification ×40 scale bars = 100 μm; magnification ×20 scale bars = 50 µm). (E) Induction of cell death in WT cells is shown in a representative analysis for annexin V staining, which demonstrated a marked increase in the percentage of apoptotic and preapoptotic cells. (F) Percentage of dead cells calculated using trypan blue staining of WT cells indicating significantly greater cell death in treated WT cells than in control cells; *p < 0.05; n = 4.

    Journal: Frontiers in Bioengineering and Biotechnology

    Article Title: A novel Frizzled 7 antibody disrupts the Wnt pathway and inhibits Wilms tumor growth

    doi: 10.3389/fbioe.2025.1641137

    Figure Lengend Snippet: The αFZD7-288.1 antibody reduces proliferation in Wilms tumors cells and induces cell death. (A) Inhibition of the Wnt pathway by αFZD7-288.1 treatment induced changes in cell kinetics, as demonstrated by the longer doubling time of antibody treated Wilms tumor (WT) cells compared to control treated WT cells and (B) the significantly lower growth rate (defined as the number of cell divisions per day), *p < 0.05; n = 4. Reduced proliferation in cells treated with αFZD7-288.1 was also observed by MTS assay in (C) and (D) Quantification of Ki67 staining was performed by counting the number of Ki67-positive nuclei in five high-power fields (HPF) per condition, using representative regions from each sample. Reduced nuclear Ki67 expression (red, lower panels) compared to that in control untreated cells (magnification ×40 scale bars = 100 μm; magnification ×20 scale bars = 50 µm). (E) Induction of cell death in WT cells is shown in a representative analysis for annexin V staining, which demonstrated a marked increase in the percentage of apoptotic and preapoptotic cells. (F) Percentage of dead cells calculated using trypan blue staining of WT cells indicating significantly greater cell death in treated WT cells than in control cells; *p < 0.05; n = 4.

    Article Snippet: The cells were stained with an isotype control (R and D Systems Cat# IC006A, RRID:AB_357254) or a primary antibody against FZD7 (R and D Systems Cat# FAB1981A, RRID:AB_2232278).

    Techniques: Inhibition, Wilms Tumor Assay, Control, MTS Assay, Staining, Expressing

    The migration and sphere formation of Wilms tumor cells is reduced after treatment with αFZD7-288.1 antibody. (A) Scratch assay demonstrating the effects of αFZD7-288.1 on the ability of cells to migrate through a scrape in the cell monolayer. Wilms tumor (WT)-treated cells demonstrated a lower migration capacity than control cells, as shown in the representative images. Calculation of the area of migrating cells (on the right) showed that the cells had a significantly reduced migration capacity following treatment; *p < 0.05; n = 3. (B) A sphere formation assay was performed. The number and size of the spheres formed by the treated cells were significantly lower than those formed by the untreated cells. The data are presented as the means ± SEMs of at least three experiments performed in triplicate (*p < 0.05) On the left, representative phase-contrast images of spheres formed by treated and untreated WT cells showing markedly fewer condensed spheres in the treated cells (scale bars = 100 μm, magnification ×10).

    Journal: Frontiers in Bioengineering and Biotechnology

    Article Title: A novel Frizzled 7 antibody disrupts the Wnt pathway and inhibits Wilms tumor growth

    doi: 10.3389/fbioe.2025.1641137

    Figure Lengend Snippet: The migration and sphere formation of Wilms tumor cells is reduced after treatment with αFZD7-288.1 antibody. (A) Scratch assay demonstrating the effects of αFZD7-288.1 on the ability of cells to migrate through a scrape in the cell monolayer. Wilms tumor (WT)-treated cells demonstrated a lower migration capacity than control cells, as shown in the representative images. Calculation of the area of migrating cells (on the right) showed that the cells had a significantly reduced migration capacity following treatment; *p < 0.05; n = 3. (B) A sphere formation assay was performed. The number and size of the spheres formed by the treated cells were significantly lower than those formed by the untreated cells. The data are presented as the means ± SEMs of at least three experiments performed in triplicate (*p < 0.05) On the left, representative phase-contrast images of spheres formed by treated and untreated WT cells showing markedly fewer condensed spheres in the treated cells (scale bars = 100 μm, magnification ×10).

    Article Snippet: The cells were stained with an isotype control (R and D Systems Cat# IC006A, RRID:AB_357254) or a primary antibody against FZD7 (R and D Systems Cat# FAB1981A, RRID:AB_2232278).

    Techniques: Migration, Wilms Tumor Assay, Wound Healing Assay, Control, Tube Formation Assay

    αFZD7-288.1 strongly co-localizes with NCAM1 in WT, and treatment with αFZD7-288.1 reduces WT CSC markers. (A) Immunofluorescence of staining of cultured WT cells using αFZD7-288.1 and NCAM1 and co-staining (αFZD7-red, αNCAM1 green and DAPI blue) microscopy revealed overlapping signals as a merged color (yellow), indicating membrane spatial proximity. Slides were imaged using Olympus IX83 fluorescent microscope and Olympus DP80 camera. Image processing was done using Olympus OlyVia software. (B) The expression of the Cancer stem cell marker genes NCAM1, CITED, SIX2 and ALDH1A1 decreased in the antibody αFZD7-288.1 treated WT derived cells (n = 3); *p < 0.05; **p < 0.01.

    Journal: Frontiers in Bioengineering and Biotechnology

    Article Title: A novel Frizzled 7 antibody disrupts the Wnt pathway and inhibits Wilms tumor growth

    doi: 10.3389/fbioe.2025.1641137

    Figure Lengend Snippet: αFZD7-288.1 strongly co-localizes with NCAM1 in WT, and treatment with αFZD7-288.1 reduces WT CSC markers. (A) Immunofluorescence of staining of cultured WT cells using αFZD7-288.1 and NCAM1 and co-staining (αFZD7-red, αNCAM1 green and DAPI blue) microscopy revealed overlapping signals as a merged color (yellow), indicating membrane spatial proximity. Slides were imaged using Olympus IX83 fluorescent microscope and Olympus DP80 camera. Image processing was done using Olympus OlyVia software. (B) The expression of the Cancer stem cell marker genes NCAM1, CITED, SIX2 and ALDH1A1 decreased in the antibody αFZD7-288.1 treated WT derived cells (n = 3); *p < 0.05; **p < 0.01.

    Article Snippet: The cells were stained with an isotype control (R and D Systems Cat# IC006A, RRID:AB_357254) or a primary antibody against FZD7 (R and D Systems Cat# FAB1981A, RRID:AB_2232278).

    Techniques: Immunofluorescence, Staining, Cell Culture, Microscopy, Membrane, Software, Expressing, Marker, Derivative Assay

    The antitumor activity of αFZD7-288.1 as demonstrated in changes in tumor volumes. (A) Flow cytometry histogram evaluating Frizzled 7 (FZD7) expression in Wilms tumor (WT) tissue for use in vivo . (B) Workflow diagram illustrating the in vivo experimental design. (C) Mouse weights during treatment. Paclitaxel (PTX)-treated mice suffered from toxic side effects and significant weight loss following treatment; **p < 0.01; ***p < 0.005. (D) Antitumor activity: Mice were treated intravenously with normal saline, αFZD7-288.1 (10 mg/kg) or paclitaxel (15 mg/kg). Tumor volume, presented in mm 3 , was assessed every 2–3 days using an external electronic caliper and calculated by the modified ellipsoid formula V = 0.52 × (length × width 2 ). Like PTX-treated tumors, tumors treated with αFZD7-288.1 exhibited growth inhibition and even tumor shrinkage, while saline-treated tumors showed rapid tumor growth; *p < 0.05; **p < 0.01. (E) Representative images on day 10 of the experiment showing mice with large saline-treated tumors (right) and unnoticeable αFZD7-288.1-treated tumors (left). (F) Waterfall graph showing the changes in the individual tumor volumes with respect to the initial tumor size. (G) FZD7 expression in tumors treated with αFZD7-288.1 was significantly altered by 0.445-fold; *p < 0.05; n = 3 per group. (H) The expression of the canonical Wnt pathway genes FZD7, β-CATENIN, AXIN2, CCND1, C-MYC, DKK1, and sFRP1 decreased in the treated tumors (n = 4); *p < 0.05; **p < 0.01; ***p < 0.005.

    Journal: Frontiers in Bioengineering and Biotechnology

    Article Title: A novel Frizzled 7 antibody disrupts the Wnt pathway and inhibits Wilms tumor growth

    doi: 10.3389/fbioe.2025.1641137

    Figure Lengend Snippet: The antitumor activity of αFZD7-288.1 as demonstrated in changes in tumor volumes. (A) Flow cytometry histogram evaluating Frizzled 7 (FZD7) expression in Wilms tumor (WT) tissue for use in vivo . (B) Workflow diagram illustrating the in vivo experimental design. (C) Mouse weights during treatment. Paclitaxel (PTX)-treated mice suffered from toxic side effects and significant weight loss following treatment; **p < 0.01; ***p < 0.005. (D) Antitumor activity: Mice were treated intravenously with normal saline, αFZD7-288.1 (10 mg/kg) or paclitaxel (15 mg/kg). Tumor volume, presented in mm 3 , was assessed every 2–3 days using an external electronic caliper and calculated by the modified ellipsoid formula V = 0.52 × (length × width 2 ). Like PTX-treated tumors, tumors treated with αFZD7-288.1 exhibited growth inhibition and even tumor shrinkage, while saline-treated tumors showed rapid tumor growth; *p < 0.05; **p < 0.01. (E) Representative images on day 10 of the experiment showing mice with large saline-treated tumors (right) and unnoticeable αFZD7-288.1-treated tumors (left). (F) Waterfall graph showing the changes in the individual tumor volumes with respect to the initial tumor size. (G) FZD7 expression in tumors treated with αFZD7-288.1 was significantly altered by 0.445-fold; *p < 0.05; n = 3 per group. (H) The expression of the canonical Wnt pathway genes FZD7, β-CATENIN, AXIN2, CCND1, C-MYC, DKK1, and sFRP1 decreased in the treated tumors (n = 4); *p < 0.05; **p < 0.01; ***p < 0.005.

    Article Snippet: The cells were stained with an isotype control (R and D Systems Cat# IC006A, RRID:AB_357254) or a primary antibody against FZD7 (R and D Systems Cat# FAB1981A, RRID:AB_2232278).

    Techniques: Activity Assay, Flow Cytometry, Expressing, Wilms Tumor Assay, In Vivo, Saline, Modification, Inhibition

    The antitumor activity of αFZD7-288.1 as demonstrated by proliferative markers in Immunohistochemical staining. (A) Immunohistochemical staining for Ki67 and Caspase3 in representative tumors from the different experimental groups; saline, paclitaxel (PTX) and αFZD7-288.1 (type A, representing reduced-volume tumors; and type B, representing tumors with growth inhibition compared to saline-treated tumors). The scale bars represent 50 μm at a magnification of ×20. Compared with those of saline- or PTX-treated tumors, αFZD7-288.1-treated tumors exhibited reduced proliferation. An increase in apoptosis was observed in αFZD7-288.1- and PTX-treated tumors (B) .

    Journal: Frontiers in Bioengineering and Biotechnology

    Article Title: A novel Frizzled 7 antibody disrupts the Wnt pathway and inhibits Wilms tumor growth

    doi: 10.3389/fbioe.2025.1641137

    Figure Lengend Snippet: The antitumor activity of αFZD7-288.1 as demonstrated by proliferative markers in Immunohistochemical staining. (A) Immunohistochemical staining for Ki67 and Caspase3 in representative tumors from the different experimental groups; saline, paclitaxel (PTX) and αFZD7-288.1 (type A, representing reduced-volume tumors; and type B, representing tumors with growth inhibition compared to saline-treated tumors). The scale bars represent 50 μm at a magnification of ×20. Compared with those of saline- or PTX-treated tumors, αFZD7-288.1-treated tumors exhibited reduced proliferation. An increase in apoptosis was observed in αFZD7-288.1- and PTX-treated tumors (B) .

    Article Snippet: The cells were stained with an isotype control (R and D Systems Cat# IC006A, RRID:AB_357254) or a primary antibody against FZD7 (R and D Systems Cat# FAB1981A, RRID:AB_2232278).

    Techniques: Activity Assay, Immunohistochemical staining, Staining, Saline, Inhibition

    ( A , C , E , G , I ) FZD7 is significantly up-regulated in GSE11691, GSE7305, GSE5108, GSE23339 and GSE25628 datasets. ( B , D , F , H , J ) The receiver-operating characteristic (ROC) curve and the area under the curve (AUC) curve of FZD7 in GSE11691, GSE7305, GSE5108, GSE23339 and GSE25628 datasets.

    Journal: Scientific Reports

    Article Title: Identification of FZD7 as a potential ferroptosis-related diagnostic gene in endometriosis by bioinformatics analysis

    doi: 10.1038/s41598-025-90803-9

    Figure Lengend Snippet: ( A , C , E , G , I ) FZD7 is significantly up-regulated in GSE11691, GSE7305, GSE5108, GSE23339 and GSE25628 datasets. ( B , D , F , H , J ) The receiver-operating characteristic (ROC) curve and the area under the curve (AUC) curve of FZD7 in GSE11691, GSE7305, GSE5108, GSE23339 and GSE25628 datasets.

    Article Snippet: The membrane was blocked with 5% skimmed milk powder solution for 2 h, then incubated with primary antibody against FZD7 (16974-1-AP, Proteintech, China) overnight at 4 °C, followed by incubation with secondary antibody for 2 h at room temperature.

    Techniques:

    GO and KEGG pathways for the overlapping FZD7 positively associated genes in GSE11691, GSE7305, GSE5108, GSE23339 and GSE25628 datasets.

    Journal: Scientific Reports

    Article Title: Identification of FZD7 as a potential ferroptosis-related diagnostic gene in endometriosis by bioinformatics analysis

    doi: 10.1038/s41598-025-90803-9

    Figure Lengend Snippet: GO and KEGG pathways for the overlapping FZD7 positively associated genes in GSE11691, GSE7305, GSE5108, GSE23339 and GSE25628 datasets.

    Article Snippet: The membrane was blocked with 5% skimmed milk powder solution for 2 h, then incubated with primary antibody against FZD7 (16974-1-AP, Proteintech, China) overnight at 4 °C, followed by incubation with secondary antibody for 2 h at room temperature.

    Techniques:

    ( A , C ) The infiltration of 22 immune cells in EMS of GSE11691dataset. ( B ) The relationship between FZD7 and 22 immune cells in GSE11691 dataset. ( D ) The expression of FZD7 in GSE19834. ( E ) Immunofluorescence images staining with CD206 in Vehicle group and F7H group.

    Journal: Scientific Reports

    Article Title: Identification of FZD7 as a potential ferroptosis-related diagnostic gene in endometriosis by bioinformatics analysis

    doi: 10.1038/s41598-025-90803-9

    Figure Lengend Snippet: ( A , C ) The infiltration of 22 immune cells in EMS of GSE11691dataset. ( B ) The relationship between FZD7 and 22 immune cells in GSE11691 dataset. ( D ) The expression of FZD7 in GSE19834. ( E ) Immunofluorescence images staining with CD206 in Vehicle group and F7H group.

    Article Snippet: The membrane was blocked with 5% skimmed milk powder solution for 2 h, then incubated with primary antibody against FZD7 (16974-1-AP, Proteintech, China) overnight at 4 °C, followed by incubation with secondary antibody for 2 h at room temperature.

    Techniques: Expressing, Immunofluorescence, Staining

    ( A-C ) The expression of FZD7 in ectopic (Ec) endometrial tissues, eutopic (Eu) endometrial tissues and normal endometrial (NE) tissues. ** p < 0.01, *** p < 0.001, ns, no significant difference by one-way analysis of variance (ANOVA) test.

    Journal: Scientific Reports

    Article Title: Identification of FZD7 as a potential ferroptosis-related diagnostic gene in endometriosis by bioinformatics analysis

    doi: 10.1038/s41598-025-90803-9

    Figure Lengend Snippet: ( A-C ) The expression of FZD7 in ectopic (Ec) endometrial tissues, eutopic (Eu) endometrial tissues and normal endometrial (NE) tissues. ** p < 0.01, *** p < 0.001, ns, no significant difference by one-way analysis of variance (ANOVA) test.

    Article Snippet: The membrane was blocked with 5% skimmed milk powder solution for 2 h, then incubated with primary antibody against FZD7 (16974-1-AP, Proteintech, China) overnight at 4 °C, followed by incubation with secondary antibody for 2 h at room temperature.

    Techniques: Expressing