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polyclonal rabbit anti goat factor b antibody  (Quidel)


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    Quidel polyclonal rabbit anti goat factor b antibody
    ( A ) Dot plot illustrating the expression of complement genes representing the classical (CP), lectin (LP) and alternative (AP) pathway in intestinal tissue biopsies from uninflamed and non-inflamed regions of patients with ulcerative colitis (UC, Single Cell Portal: SCP259). ( B ) Plots depict CFB expression across denoted colonic cell types in the same dataset from intestinal tissue of patients with UC and healthy controls. * P <0.05, *** P <0.001, ns, not significant, Wilcoxon rank test. ( C ) Immunofluorescence staining for <t>Factor</t> <t>B</t> (red), E-cadherin (green) and DAPI (blue) in colonic tissue specimens from patients with IBD versus controls (non-IBD). (Scale bar = 90 μ m). ( D ) Quantification of the relative intensity of Factor B expression across tissue type. ( E ) Representative spatial transcriptomic slides (Visium) showing CFB expression in intestinal tissues obtained from patients with UC compared to healthy controls (HC, GSE189184). Graph depicts quantification of CFB expression from 3 HC and 7 UC specimens. ( F ) Representative spatial transcriptomic slides showing DUOX2-CFB co-localization spots in HC (left) and UC tissue (right) from the same dataset. Bar plot represents average co-expression across tissue samples. Error bars represent standard error of the mean (SEM). * P < 0.05, unpaired t-test.
    Polyclonal Rabbit Anti Goat Factor B Antibody, supplied by Quidel, used in various techniques. Bioz Stars score: 95/100, based on 37 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/polyclonal rabbit anti goat factor b antibody/product/Quidel
    Average 95 stars, based on 37 article reviews
    polyclonal rabbit anti goat factor b antibody - by Bioz Stars, 2026-05
    95/100 stars

    Images

    1) Product Images from "Mucosally sourced complement factor B modulates the host response to colitis"

    Article Title: Mucosally sourced complement factor B modulates the host response to colitis

    Journal: bioRxiv

    doi: 10.64898/2025.12.31.697141

    ( A ) Dot plot illustrating the expression of complement genes representing the classical (CP), lectin (LP) and alternative (AP) pathway in intestinal tissue biopsies from uninflamed and non-inflamed regions of patients with ulcerative colitis (UC, Single Cell Portal: SCP259). ( B ) Plots depict CFB expression across denoted colonic cell types in the same dataset from intestinal tissue of patients with UC and healthy controls. * P <0.05, *** P <0.001, ns, not significant, Wilcoxon rank test. ( C ) Immunofluorescence staining for Factor B (red), E-cadherin (green) and DAPI (blue) in colonic tissue specimens from patients with IBD versus controls (non-IBD). (Scale bar = 90 μ m). ( D ) Quantification of the relative intensity of Factor B expression across tissue type. ( E ) Representative spatial transcriptomic slides (Visium) showing CFB expression in intestinal tissues obtained from patients with UC compared to healthy controls (HC, GSE189184). Graph depicts quantification of CFB expression from 3 HC and 7 UC specimens. ( F ) Representative spatial transcriptomic slides showing DUOX2-CFB co-localization spots in HC (left) and UC tissue (right) from the same dataset. Bar plot represents average co-expression across tissue samples. Error bars represent standard error of the mean (SEM). * P < 0.05, unpaired t-test.
    Figure Legend Snippet: ( A ) Dot plot illustrating the expression of complement genes representing the classical (CP), lectin (LP) and alternative (AP) pathway in intestinal tissue biopsies from uninflamed and non-inflamed regions of patients with ulcerative colitis (UC, Single Cell Portal: SCP259). ( B ) Plots depict CFB expression across denoted colonic cell types in the same dataset from intestinal tissue of patients with UC and healthy controls. * P <0.05, *** P <0.001, ns, not significant, Wilcoxon rank test. ( C ) Immunofluorescence staining for Factor B (red), E-cadherin (green) and DAPI (blue) in colonic tissue specimens from patients with IBD versus controls (non-IBD). (Scale bar = 90 μ m). ( D ) Quantification of the relative intensity of Factor B expression across tissue type. ( E ) Representative spatial transcriptomic slides (Visium) showing CFB expression in intestinal tissues obtained from patients with UC compared to healthy controls (HC, GSE189184). Graph depicts quantification of CFB expression from 3 HC and 7 UC specimens. ( F ) Representative spatial transcriptomic slides showing DUOX2-CFB co-localization spots in HC (left) and UC tissue (right) from the same dataset. Bar plot represents average co-expression across tissue samples. Error bars represent standard error of the mean (SEM). * P < 0.05, unpaired t-test.

    Techniques Used: Expressing, Immunofluorescence, Staining

    ( A ) Immunoblot denoting Factor B (FB) in plasma of global Cfb knockouts ( Cfb −/− ), mice deficient in liver-derived FB ( Cfb f/f AlbCre +/− ) and their littermates ( Cfb f/f AlbCre −/− ). MM: molecular marker. The same volume of plasma was loaded per well at 1:20 dilution. ( B ) Cfb expression on qRT-PCR in the colons from indicated mice, values normalized to Hprt1 . ( C ) Quantification of the relative intensity of Factor B expression in colon tissue from indicated mice. ( D ) Representative immunofluorescence images of colonic tissue from Cfb −/− , Cfb f/f AlbCre +/− and Cfb f/ f AlbCre −/− mice (n=4/group), stained with Factor B (red), E-cadherin (green) and DAPI (blue). Scale bars = 70 μ m. ( E ) Graph depicts serum CD14 concentration. Serum from mice challenged with lipopolysaccharide (LPS) were used as a positive control for barrier leak. Each dot represents an individual mouse. Bar graphs show the mean ± SEM, * P < 0.05; ** P < 0.01, ns, non-significant, unpaired t test. ( F ) Dot plot illustrating the expression of complement genes representing the classical (CP), lectin (LP) and alternative (AP) pathway across cell types indicated along the Y axis, based on single-cell RNA-seq data from the large intestines of non-injured adult C57BL/6 mice ( Cfb f/f ), housed in specific pathogen-free conditions. Cells pooled from n=2 mice.
    Figure Legend Snippet: ( A ) Immunoblot denoting Factor B (FB) in plasma of global Cfb knockouts ( Cfb −/− ), mice deficient in liver-derived FB ( Cfb f/f AlbCre +/− ) and their littermates ( Cfb f/f AlbCre −/− ). MM: molecular marker. The same volume of plasma was loaded per well at 1:20 dilution. ( B ) Cfb expression on qRT-PCR in the colons from indicated mice, values normalized to Hprt1 . ( C ) Quantification of the relative intensity of Factor B expression in colon tissue from indicated mice. ( D ) Representative immunofluorescence images of colonic tissue from Cfb −/− , Cfb f/f AlbCre +/− and Cfb f/ f AlbCre −/− mice (n=4/group), stained with Factor B (red), E-cadherin (green) and DAPI (blue). Scale bars = 70 μ m. ( E ) Graph depicts serum CD14 concentration. Serum from mice challenged with lipopolysaccharide (LPS) were used as a positive control for barrier leak. Each dot represents an individual mouse. Bar graphs show the mean ± SEM, * P < 0.05; ** P < 0.01, ns, non-significant, unpaired t test. ( F ) Dot plot illustrating the expression of complement genes representing the classical (CP), lectin (LP) and alternative (AP) pathway across cell types indicated along the Y axis, based on single-cell RNA-seq data from the large intestines of non-injured adult C57BL/6 mice ( Cfb f/f ), housed in specific pathogen-free conditions. Cells pooled from n=2 mice.

    Techniques Used: Western Blot, Clinical Proteomics, Derivative Assay, Marker, Expressing, Quantitative RT-PCR, Immunofluorescence, Staining, Concentration Assay, Positive Control, RNA Sequencing

    ( A-B ) Human epithelial Caco-2 cells were incubated in serum-free media alone (non-treated, NT), media with rIL-1β (50ng/ml) or TNF-α (50ng/ml). ( A ) Supernatant was collected after 24 hours, concentrated 50-fold and analyzed by immunoblotting for Factor B. ( B ) Human primary fibroblasts were incubated in serum-free media alone (NT), media with rIL-1β (50ng/ml) or TNF-α (50ng/ml). Immunoblot depicts CFB in supernatant at the 24 h time point. Graph depicts Factor B protein quantification relative to the loaded control. The same volume of supernatant was loaded in each well. Each dot represents an individual replicate. Bar graphs show the mean ± SEM. All immunoblots were repeated at least twice. Statistics performed using an ordinary one-way ANOVA test adjusted for multiple comparisons.
    Figure Legend Snippet: ( A-B ) Human epithelial Caco-2 cells were incubated in serum-free media alone (non-treated, NT), media with rIL-1β (50ng/ml) or TNF-α (50ng/ml). ( A ) Supernatant was collected after 24 hours, concentrated 50-fold and analyzed by immunoblotting for Factor B. ( B ) Human primary fibroblasts were incubated in serum-free media alone (NT), media with rIL-1β (50ng/ml) or TNF-α (50ng/ml). Immunoblot depicts CFB in supernatant at the 24 h time point. Graph depicts Factor B protein quantification relative to the loaded control. The same volume of supernatant was loaded in each well. Each dot represents an individual replicate. Bar graphs show the mean ± SEM. All immunoblots were repeated at least twice. Statistics performed using an ordinary one-way ANOVA test adjusted for multiple comparisons.

    Techniques Used: Incubation, Western Blot, Control

    ( A ) SPF-housed global knockout ( Cfb −/− ), liver-deficient ( Cfb f/f AlbCre +/− ) and littermate control ( Cfb f/f AlbCre −/− ) mice were administered 2.5% DSS in drinking water for 7 days, followed by conventional water for 3 days. Mice were euthanized on Day 11. Body weights were monitored daily during treatment. ( B ) Colon lengths evaluated at end of study. ( C ) Colonic tissue was homogenized for measuring TNF-α and MPO levels, normalized to weight (per g) of tissue. ( D ) Fecal lipocalin-2 (LCN-2) levels compared at the end of the study. ( E ) Complement activity in the colonic tissue measured using C3bBbP (alternative pathway convertase). ( F ) Differentially regulated pathways upregulated in Cfb −/− versus Cfb f/f AlbCre +/− based on bulk RNA-seq of colonic tissue at end of treatment. Pathway analysis conducted using Enrichr (MSigDB). ( G ) Adult C57BL/6 mice housed in SPF conditions were given 2.5% DSS in drinking water for a week. Starting day 5, mice were treated with a daily oral gavage of a Factor B inhibitor, iptacopan (20 mg/kg), or vehicle control. Body weights were monitored daily. ( H ) Colon lengths compared at end of the experiment. ( I ) IL-1β in colonic tissue lysate and ( J ) LCN-2 in fecal samples. Bar graphs show the mean ± SEM, * P < 0.05; ** P < 0.01, ns, non-significant. Unpaired t test when analyzing a two-group categorical comparison, and 2-way ANOVA adjusted for multiple comparisons when analyzing longitudinal data.
    Figure Legend Snippet: ( A ) SPF-housed global knockout ( Cfb −/− ), liver-deficient ( Cfb f/f AlbCre +/− ) and littermate control ( Cfb f/f AlbCre −/− ) mice were administered 2.5% DSS in drinking water for 7 days, followed by conventional water for 3 days. Mice were euthanized on Day 11. Body weights were monitored daily during treatment. ( B ) Colon lengths evaluated at end of study. ( C ) Colonic tissue was homogenized for measuring TNF-α and MPO levels, normalized to weight (per g) of tissue. ( D ) Fecal lipocalin-2 (LCN-2) levels compared at the end of the study. ( E ) Complement activity in the colonic tissue measured using C3bBbP (alternative pathway convertase). ( F ) Differentially regulated pathways upregulated in Cfb −/− versus Cfb f/f AlbCre +/− based on bulk RNA-seq of colonic tissue at end of treatment. Pathway analysis conducted using Enrichr (MSigDB). ( G ) Adult C57BL/6 mice housed in SPF conditions were given 2.5% DSS in drinking water for a week. Starting day 5, mice were treated with a daily oral gavage of a Factor B inhibitor, iptacopan (20 mg/kg), or vehicle control. Body weights were monitored daily. ( H ) Colon lengths compared at end of the experiment. ( I ) IL-1β in colonic tissue lysate and ( J ) LCN-2 in fecal samples. Bar graphs show the mean ± SEM, * P < 0.05; ** P < 0.01, ns, non-significant. Unpaired t test when analyzing a two-group categorical comparison, and 2-way ANOVA adjusted for multiple comparisons when analyzing longitudinal data.

    Techniques Used: Knock-Out, Control, Activity Assay, RNA Sequencing, Comparison

    ( A-C ) Data from scRNA-seq of colonic cells from C57BL/6 mice subjected to acute DSS (acute colitis, AC) was re-analyzed for express of complement genes and compared to mice not treated with DSS (non-DSS, healthy controls (HC), GSE264408). ( A ) UMAP with designated cell clusters showing Cfb expression. ( B ) Bar plots depicting changes in the cellular composition of the colonic tissue post-DSS treatment. ( C ) Dot plots showing average gene expression levels of key AP components in the distinct cell clusters of HC (healthy control) mice and those with AC (acute colitis). ( D ) Cfb f/f LysMCre +/− mice and controls were treated with 7 days of 2.5% DSS in drinking water, followed by 3 days of regular water. Body weights were monitored daily. Colon length was determined at the end of the experiment. Colonic tissue was homogenized for comparing MPO and IL-1β levels, normalized to tissue weight. ( E ) Representative RNAScope images depict staining of colonic tissue from Cfb f/f VilCre +/− mice and controls. Scale bar =30μm. ( F ) Cfb f/f VilCre and controls were treated similarly to ( D ). Body weights and the disease activity index (DAI) were measured daily. Mice were sacrificed on day 11. Colons were harvested to compare the colon length, MPO and IL-1β levels, normalized to weight of tissue. ( G ) Cfb f/f Col1a2CreER T2+/− mice and their controls were treated with tamoxifen every other day for 10 days. Colons were harvested and tissue sections were stained for Factor B (red) and vimentin (green). Scale bar =90 μ m. ( H ) Cfb f/f Col1a2CreER T2+/− and littermates were subjected to tamoxifen challenge as detailed in ( G ). 3 days after the last dose of tamoxifen, mice were subjected to DSS challenge as detailed in ( D ). Body weights and DAI scores were monitored daily. Colon tissue was collected at day 11 to measure colon length and cytokines in tissue lysate. Data represent 2 independent experiments. Bar graphs show the mean ± SEM, * P < 0.05; ** P < 0.01, ns, non-significant. Unpaired t test when analyzing a two-group categorical comparison, and 2-way ANOVA adjusted for multiple comparisons when analyzing longitudinal data.
    Figure Legend Snippet: ( A-C ) Data from scRNA-seq of colonic cells from C57BL/6 mice subjected to acute DSS (acute colitis, AC) was re-analyzed for express of complement genes and compared to mice not treated with DSS (non-DSS, healthy controls (HC), GSE264408). ( A ) UMAP with designated cell clusters showing Cfb expression. ( B ) Bar plots depicting changes in the cellular composition of the colonic tissue post-DSS treatment. ( C ) Dot plots showing average gene expression levels of key AP components in the distinct cell clusters of HC (healthy control) mice and those with AC (acute colitis). ( D ) Cfb f/f LysMCre +/− mice and controls were treated with 7 days of 2.5% DSS in drinking water, followed by 3 days of regular water. Body weights were monitored daily. Colon length was determined at the end of the experiment. Colonic tissue was homogenized for comparing MPO and IL-1β levels, normalized to tissue weight. ( E ) Representative RNAScope images depict staining of colonic tissue from Cfb f/f VilCre +/− mice and controls. Scale bar =30μm. ( F ) Cfb f/f VilCre and controls were treated similarly to ( D ). Body weights and the disease activity index (DAI) were measured daily. Mice were sacrificed on day 11. Colons were harvested to compare the colon length, MPO and IL-1β levels, normalized to weight of tissue. ( G ) Cfb f/f Col1a2CreER T2+/− mice and their controls were treated with tamoxifen every other day for 10 days. Colons were harvested and tissue sections were stained for Factor B (red) and vimentin (green). Scale bar =90 μ m. ( H ) Cfb f/f Col1a2CreER T2+/− and littermates were subjected to tamoxifen challenge as detailed in ( G ). 3 days after the last dose of tamoxifen, mice were subjected to DSS challenge as detailed in ( D ). Body weights and DAI scores were monitored daily. Colon tissue was collected at day 11 to measure colon length and cytokines in tissue lysate. Data represent 2 independent experiments. Bar graphs show the mean ± SEM, * P < 0.05; ** P < 0.01, ns, non-significant. Unpaired t test when analyzing a two-group categorical comparison, and 2-way ANOVA adjusted for multiple comparisons when analyzing longitudinal data.

    Techniques Used: Expressing, Gene Expression, Control, RNAscope, Staining, Activity Assay, Comparison



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    Image Search Results


    ( A ) Dot plot illustrating the expression of complement genes representing the classical (CP), lectin (LP) and alternative (AP) pathway in intestinal tissue biopsies from uninflamed and non-inflamed regions of patients with ulcerative colitis (UC, Single Cell Portal: SCP259). ( B ) Plots depict CFB expression across denoted colonic cell types in the same dataset from intestinal tissue of patients with UC and healthy controls. * P <0.05, *** P <0.001, ns, not significant, Wilcoxon rank test. ( C ) Immunofluorescence staining for Factor B (red), E-cadherin (green) and DAPI (blue) in colonic tissue specimens from patients with IBD versus controls (non-IBD). (Scale bar = 90 μ m). ( D ) Quantification of the relative intensity of Factor B expression across tissue type. ( E ) Representative spatial transcriptomic slides (Visium) showing CFB expression in intestinal tissues obtained from patients with UC compared to healthy controls (HC, GSE189184). Graph depicts quantification of CFB expression from 3 HC and 7 UC specimens. ( F ) Representative spatial transcriptomic slides showing DUOX2-CFB co-localization spots in HC (left) and UC tissue (right) from the same dataset. Bar plot represents average co-expression across tissue samples. Error bars represent standard error of the mean (SEM). * P < 0.05, unpaired t-test.

    Journal: bioRxiv

    Article Title: Mucosally sourced complement factor B modulates the host response to colitis

    doi: 10.64898/2025.12.31.697141

    Figure Lengend Snippet: ( A ) Dot plot illustrating the expression of complement genes representing the classical (CP), lectin (LP) and alternative (AP) pathway in intestinal tissue biopsies from uninflamed and non-inflamed regions of patients with ulcerative colitis (UC, Single Cell Portal: SCP259). ( B ) Plots depict CFB expression across denoted colonic cell types in the same dataset from intestinal tissue of patients with UC and healthy controls. * P <0.05, *** P <0.001, ns, not significant, Wilcoxon rank test. ( C ) Immunofluorescence staining for Factor B (red), E-cadherin (green) and DAPI (blue) in colonic tissue specimens from patients with IBD versus controls (non-IBD). (Scale bar = 90 μ m). ( D ) Quantification of the relative intensity of Factor B expression across tissue type. ( E ) Representative spatial transcriptomic slides (Visium) showing CFB expression in intestinal tissues obtained from patients with UC compared to healthy controls (HC, GSE189184). Graph depicts quantification of CFB expression from 3 HC and 7 UC specimens. ( F ) Representative spatial transcriptomic slides showing DUOX2-CFB co-localization spots in HC (left) and UC tissue (right) from the same dataset. Bar plot represents average co-expression across tissue samples. Error bars represent standard error of the mean (SEM). * P < 0.05, unpaired t-test.

    Article Snippet: The sections were then blocked with 3% Bovine Albumen (Thermo Fisher, #J10857.22) diluted in phosphate buffered saline (PBS) at room temperature for 1 h. For primary incubation the sections were stained with a 1:50 dilution of polyclonal rabbit anti-goat Factor B antibody (Quidel Ortho, # A311) and a 1:100 dilution of mouse anti-E-Cadherin (BD Sciences, Cat. 610181) overnight at 4°C.

    Techniques: Expressing, Immunofluorescence, Staining

    ( A ) Immunoblot denoting Factor B (FB) in plasma of global Cfb knockouts ( Cfb −/− ), mice deficient in liver-derived FB ( Cfb f/f AlbCre +/− ) and their littermates ( Cfb f/f AlbCre −/− ). MM: molecular marker. The same volume of plasma was loaded per well at 1:20 dilution. ( B ) Cfb expression on qRT-PCR in the colons from indicated mice, values normalized to Hprt1 . ( C ) Quantification of the relative intensity of Factor B expression in colon tissue from indicated mice. ( D ) Representative immunofluorescence images of colonic tissue from Cfb −/− , Cfb f/f AlbCre +/− and Cfb f/ f AlbCre −/− mice (n=4/group), stained with Factor B (red), E-cadherin (green) and DAPI (blue). Scale bars = 70 μ m. ( E ) Graph depicts serum CD14 concentration. Serum from mice challenged with lipopolysaccharide (LPS) were used as a positive control for barrier leak. Each dot represents an individual mouse. Bar graphs show the mean ± SEM, * P < 0.05; ** P < 0.01, ns, non-significant, unpaired t test. ( F ) Dot plot illustrating the expression of complement genes representing the classical (CP), lectin (LP) and alternative (AP) pathway across cell types indicated along the Y axis, based on single-cell RNA-seq data from the large intestines of non-injured adult C57BL/6 mice ( Cfb f/f ), housed in specific pathogen-free conditions. Cells pooled from n=2 mice.

    Journal: bioRxiv

    Article Title: Mucosally sourced complement factor B modulates the host response to colitis

    doi: 10.64898/2025.12.31.697141

    Figure Lengend Snippet: ( A ) Immunoblot denoting Factor B (FB) in plasma of global Cfb knockouts ( Cfb −/− ), mice deficient in liver-derived FB ( Cfb f/f AlbCre +/− ) and their littermates ( Cfb f/f AlbCre −/− ). MM: molecular marker. The same volume of plasma was loaded per well at 1:20 dilution. ( B ) Cfb expression on qRT-PCR in the colons from indicated mice, values normalized to Hprt1 . ( C ) Quantification of the relative intensity of Factor B expression in colon tissue from indicated mice. ( D ) Representative immunofluorescence images of colonic tissue from Cfb −/− , Cfb f/f AlbCre +/− and Cfb f/ f AlbCre −/− mice (n=4/group), stained with Factor B (red), E-cadherin (green) and DAPI (blue). Scale bars = 70 μ m. ( E ) Graph depicts serum CD14 concentration. Serum from mice challenged with lipopolysaccharide (LPS) were used as a positive control for barrier leak. Each dot represents an individual mouse. Bar graphs show the mean ± SEM, * P < 0.05; ** P < 0.01, ns, non-significant, unpaired t test. ( F ) Dot plot illustrating the expression of complement genes representing the classical (CP), lectin (LP) and alternative (AP) pathway across cell types indicated along the Y axis, based on single-cell RNA-seq data from the large intestines of non-injured adult C57BL/6 mice ( Cfb f/f ), housed in specific pathogen-free conditions. Cells pooled from n=2 mice.

    Article Snippet: The sections were then blocked with 3% Bovine Albumen (Thermo Fisher, #J10857.22) diluted in phosphate buffered saline (PBS) at room temperature for 1 h. For primary incubation the sections were stained with a 1:50 dilution of polyclonal rabbit anti-goat Factor B antibody (Quidel Ortho, # A311) and a 1:100 dilution of mouse anti-E-Cadherin (BD Sciences, Cat. 610181) overnight at 4°C.

    Techniques: Western Blot, Clinical Proteomics, Derivative Assay, Marker, Expressing, Quantitative RT-PCR, Immunofluorescence, Staining, Concentration Assay, Positive Control, RNA Sequencing

    ( A-B ) Human epithelial Caco-2 cells were incubated in serum-free media alone (non-treated, NT), media with rIL-1β (50ng/ml) or TNF-α (50ng/ml). ( A ) Supernatant was collected after 24 hours, concentrated 50-fold and analyzed by immunoblotting for Factor B. ( B ) Human primary fibroblasts were incubated in serum-free media alone (NT), media with rIL-1β (50ng/ml) or TNF-α (50ng/ml). Immunoblot depicts CFB in supernatant at the 24 h time point. Graph depicts Factor B protein quantification relative to the loaded control. The same volume of supernatant was loaded in each well. Each dot represents an individual replicate. Bar graphs show the mean ± SEM. All immunoblots were repeated at least twice. Statistics performed using an ordinary one-way ANOVA test adjusted for multiple comparisons.

    Journal: bioRxiv

    Article Title: Mucosally sourced complement factor B modulates the host response to colitis

    doi: 10.64898/2025.12.31.697141

    Figure Lengend Snippet: ( A-B ) Human epithelial Caco-2 cells were incubated in serum-free media alone (non-treated, NT), media with rIL-1β (50ng/ml) or TNF-α (50ng/ml). ( A ) Supernatant was collected after 24 hours, concentrated 50-fold and analyzed by immunoblotting for Factor B. ( B ) Human primary fibroblasts were incubated in serum-free media alone (NT), media with rIL-1β (50ng/ml) or TNF-α (50ng/ml). Immunoblot depicts CFB in supernatant at the 24 h time point. Graph depicts Factor B protein quantification relative to the loaded control. The same volume of supernatant was loaded in each well. Each dot represents an individual replicate. Bar graphs show the mean ± SEM. All immunoblots were repeated at least twice. Statistics performed using an ordinary one-way ANOVA test adjusted for multiple comparisons.

    Article Snippet: The sections were then blocked with 3% Bovine Albumen (Thermo Fisher, #J10857.22) diluted in phosphate buffered saline (PBS) at room temperature for 1 h. For primary incubation the sections were stained with a 1:50 dilution of polyclonal rabbit anti-goat Factor B antibody (Quidel Ortho, # A311) and a 1:100 dilution of mouse anti-E-Cadherin (BD Sciences, Cat. 610181) overnight at 4°C.

    Techniques: Incubation, Western Blot, Control

    ( A ) SPF-housed global knockout ( Cfb −/− ), liver-deficient ( Cfb f/f AlbCre +/− ) and littermate control ( Cfb f/f AlbCre −/− ) mice were administered 2.5% DSS in drinking water for 7 days, followed by conventional water for 3 days. Mice were euthanized on Day 11. Body weights were monitored daily during treatment. ( B ) Colon lengths evaluated at end of study. ( C ) Colonic tissue was homogenized for measuring TNF-α and MPO levels, normalized to weight (per g) of tissue. ( D ) Fecal lipocalin-2 (LCN-2) levels compared at the end of the study. ( E ) Complement activity in the colonic tissue measured using C3bBbP (alternative pathway convertase). ( F ) Differentially regulated pathways upregulated in Cfb −/− versus Cfb f/f AlbCre +/− based on bulk RNA-seq of colonic tissue at end of treatment. Pathway analysis conducted using Enrichr (MSigDB). ( G ) Adult C57BL/6 mice housed in SPF conditions were given 2.5% DSS in drinking water for a week. Starting day 5, mice were treated with a daily oral gavage of a Factor B inhibitor, iptacopan (20 mg/kg), or vehicle control. Body weights were monitored daily. ( H ) Colon lengths compared at end of the experiment. ( I ) IL-1β in colonic tissue lysate and ( J ) LCN-2 in fecal samples. Bar graphs show the mean ± SEM, * P < 0.05; ** P < 0.01, ns, non-significant. Unpaired t test when analyzing a two-group categorical comparison, and 2-way ANOVA adjusted for multiple comparisons when analyzing longitudinal data.

    Journal: bioRxiv

    Article Title: Mucosally sourced complement factor B modulates the host response to colitis

    doi: 10.64898/2025.12.31.697141

    Figure Lengend Snippet: ( A ) SPF-housed global knockout ( Cfb −/− ), liver-deficient ( Cfb f/f AlbCre +/− ) and littermate control ( Cfb f/f AlbCre −/− ) mice were administered 2.5% DSS in drinking water for 7 days, followed by conventional water for 3 days. Mice were euthanized on Day 11. Body weights were monitored daily during treatment. ( B ) Colon lengths evaluated at end of study. ( C ) Colonic tissue was homogenized for measuring TNF-α and MPO levels, normalized to weight (per g) of tissue. ( D ) Fecal lipocalin-2 (LCN-2) levels compared at the end of the study. ( E ) Complement activity in the colonic tissue measured using C3bBbP (alternative pathway convertase). ( F ) Differentially regulated pathways upregulated in Cfb −/− versus Cfb f/f AlbCre +/− based on bulk RNA-seq of colonic tissue at end of treatment. Pathway analysis conducted using Enrichr (MSigDB). ( G ) Adult C57BL/6 mice housed in SPF conditions were given 2.5% DSS in drinking water for a week. Starting day 5, mice were treated with a daily oral gavage of a Factor B inhibitor, iptacopan (20 mg/kg), or vehicle control. Body weights were monitored daily. ( H ) Colon lengths compared at end of the experiment. ( I ) IL-1β in colonic tissue lysate and ( J ) LCN-2 in fecal samples. Bar graphs show the mean ± SEM, * P < 0.05; ** P < 0.01, ns, non-significant. Unpaired t test when analyzing a two-group categorical comparison, and 2-way ANOVA adjusted for multiple comparisons when analyzing longitudinal data.

    Article Snippet: The sections were then blocked with 3% Bovine Albumen (Thermo Fisher, #J10857.22) diluted in phosphate buffered saline (PBS) at room temperature for 1 h. For primary incubation the sections were stained with a 1:50 dilution of polyclonal rabbit anti-goat Factor B antibody (Quidel Ortho, # A311) and a 1:100 dilution of mouse anti-E-Cadherin (BD Sciences, Cat. 610181) overnight at 4°C.

    Techniques: Knock-Out, Control, Activity Assay, RNA Sequencing, Comparison

    ( A-C ) Data from scRNA-seq of colonic cells from C57BL/6 mice subjected to acute DSS (acute colitis, AC) was re-analyzed for express of complement genes and compared to mice not treated with DSS (non-DSS, healthy controls (HC), GSE264408). ( A ) UMAP with designated cell clusters showing Cfb expression. ( B ) Bar plots depicting changes in the cellular composition of the colonic tissue post-DSS treatment. ( C ) Dot plots showing average gene expression levels of key AP components in the distinct cell clusters of HC (healthy control) mice and those with AC (acute colitis). ( D ) Cfb f/f LysMCre +/− mice and controls were treated with 7 days of 2.5% DSS in drinking water, followed by 3 days of regular water. Body weights were monitored daily. Colon length was determined at the end of the experiment. Colonic tissue was homogenized for comparing MPO and IL-1β levels, normalized to tissue weight. ( E ) Representative RNAScope images depict staining of colonic tissue from Cfb f/f VilCre +/− mice and controls. Scale bar =30μm. ( F ) Cfb f/f VilCre and controls were treated similarly to ( D ). Body weights and the disease activity index (DAI) were measured daily. Mice were sacrificed on day 11. Colons were harvested to compare the colon length, MPO and IL-1β levels, normalized to weight of tissue. ( G ) Cfb f/f Col1a2CreER T2+/− mice and their controls were treated with tamoxifen every other day for 10 days. Colons were harvested and tissue sections were stained for Factor B (red) and vimentin (green). Scale bar =90 μ m. ( H ) Cfb f/f Col1a2CreER T2+/− and littermates were subjected to tamoxifen challenge as detailed in ( G ). 3 days after the last dose of tamoxifen, mice were subjected to DSS challenge as detailed in ( D ). Body weights and DAI scores were monitored daily. Colon tissue was collected at day 11 to measure colon length and cytokines in tissue lysate. Data represent 2 independent experiments. Bar graphs show the mean ± SEM, * P < 0.05; ** P < 0.01, ns, non-significant. Unpaired t test when analyzing a two-group categorical comparison, and 2-way ANOVA adjusted for multiple comparisons when analyzing longitudinal data.

    Journal: bioRxiv

    Article Title: Mucosally sourced complement factor B modulates the host response to colitis

    doi: 10.64898/2025.12.31.697141

    Figure Lengend Snippet: ( A-C ) Data from scRNA-seq of colonic cells from C57BL/6 mice subjected to acute DSS (acute colitis, AC) was re-analyzed for express of complement genes and compared to mice not treated with DSS (non-DSS, healthy controls (HC), GSE264408). ( A ) UMAP with designated cell clusters showing Cfb expression. ( B ) Bar plots depicting changes in the cellular composition of the colonic tissue post-DSS treatment. ( C ) Dot plots showing average gene expression levels of key AP components in the distinct cell clusters of HC (healthy control) mice and those with AC (acute colitis). ( D ) Cfb f/f LysMCre +/− mice and controls were treated with 7 days of 2.5% DSS in drinking water, followed by 3 days of regular water. Body weights were monitored daily. Colon length was determined at the end of the experiment. Colonic tissue was homogenized for comparing MPO and IL-1β levels, normalized to tissue weight. ( E ) Representative RNAScope images depict staining of colonic tissue from Cfb f/f VilCre +/− mice and controls. Scale bar =30μm. ( F ) Cfb f/f VilCre and controls were treated similarly to ( D ). Body weights and the disease activity index (DAI) were measured daily. Mice were sacrificed on day 11. Colons were harvested to compare the colon length, MPO and IL-1β levels, normalized to weight of tissue. ( G ) Cfb f/f Col1a2CreER T2+/− mice and their controls were treated with tamoxifen every other day for 10 days. Colons were harvested and tissue sections were stained for Factor B (red) and vimentin (green). Scale bar =90 μ m. ( H ) Cfb f/f Col1a2CreER T2+/− and littermates were subjected to tamoxifen challenge as detailed in ( G ). 3 days after the last dose of tamoxifen, mice were subjected to DSS challenge as detailed in ( D ). Body weights and DAI scores were monitored daily. Colon tissue was collected at day 11 to measure colon length and cytokines in tissue lysate. Data represent 2 independent experiments. Bar graphs show the mean ± SEM, * P < 0.05; ** P < 0.01, ns, non-significant. Unpaired t test when analyzing a two-group categorical comparison, and 2-way ANOVA adjusted for multiple comparisons when analyzing longitudinal data.

    Article Snippet: The sections were then blocked with 3% Bovine Albumen (Thermo Fisher, #J10857.22) diluted in phosphate buffered saline (PBS) at room temperature for 1 h. For primary incubation the sections were stained with a 1:50 dilution of polyclonal rabbit anti-goat Factor B antibody (Quidel Ortho, # A311) and a 1:100 dilution of mouse anti-E-Cadherin (BD Sciences, Cat. 610181) overnight at 4°C.

    Techniques: Expressing, Gene Expression, Control, RNAscope, Staining, Activity Assay, Comparison

    Figure 5 Protein content of TGF-β1 (25 Kd) in SMGs of mice at 8 hours, 24 hours, and 4 weeks after IR was assessed by Western blot analysis. Notes: (A) Representative Western blots from three to five experiments with similar results. (B) Changes in TGF-β1 quantified by scanning densitometry analysis using Image Lab software. The data (relative density normalized to β-actin) are expressed as mean ± SD; *P,0.05, **P,0.01. Abbreviations: TGF-β1, transforming growth factor β1; SMG, submandibular gland; IR, irradiation; SD, standard deviation; h, hours; w, weeks.

    Journal: Drug Design, Development and Therapy

    Article Title: Simvastatin attenuates radiation-induced salivary gland dysfunction in mice

    doi: 10.2147/dddt.s105809

    Figure Lengend Snippet: Figure 5 Protein content of TGF-β1 (25 Kd) in SMGs of mice at 8 hours, 24 hours, and 4 weeks after IR was assessed by Western blot analysis. Notes: (A) Representative Western blots from three to five experiments with similar results. (B) Changes in TGF-β1 quantified by scanning densitometry analysis using Image Lab software. The data (relative density normalized to β-actin) are expressed as mean ± SD; *P,0.05, **P,0.01. Abbreviations: TGF-β1, transforming growth factor β1; SMG, submandibular gland; IR, irradiation; SD, standard deviation; h, hours; w, weeks.

    Article Snippet: The membranes were blocked in a blocking buffer (5% nonfat milk, 1% Tween 20, in 20 mM Tris-buffered saline, pH 7.6) for 1 hour at room temperature, followed by incubation with the transforming growth factor (TGF)-β1 rabbit polyclonal immunoglobulin G (IgG) (Santa Cruz Biotechnology Inc., Dallas, TX, USA) in a blocking buffer overnight at 4°C.

    Techniques: Western Blot, Software, Irradiation, Standard Deviation

    FIG. 4. Two-dimensional electrophoresis results and validation with Western blotting. A, 2DE maps of proteins extracted from control and biopsied brains. B, magnified comparison maps of spots for ENO2, Hsd17b10, UCHL1, ,-SNAP, GMFB, PRDX5, and MBP in the 2DE patterns of control and biopsied brains. Bars represent the relative volume (%Vol) of each of the spots on the gels. Error bars represent the standard error of the mean. Western blot analyses of each protein are shown simultaneously. Results are consistent with the results of the 2DE. Actin was used as a loading control.

    Journal: Molecular & Cellular Proteomics

    Article Title: Evaluation of Blastomere Biopsy Using a Mouse Model Indicates the Potential High Risk of Neurodegenerative Disorders in the Offspring

    doi: 10.1074/mcp.m800273-mcp200

    Figure Lengend Snippet: FIG. 4. Two-dimensional electrophoresis results and validation with Western blotting. A, 2DE maps of proteins extracted from control and biopsied brains. B, magnified comparison maps of spots for ENO2, Hsd17b10, UCHL1, ,-SNAP, GMFB, PRDX5, and MBP in the 2DE patterns of control and biopsied brains. Bars represent the relative volume (%Vol) of each of the spots on the gels. Error bars represent the standard error of the mean. Western blot analyses of each protein are shown simultaneously. Results are consistent with the results of the 2DE. Actin was used as a loading control.

    Article Snippet: Primary antibodies used were anti-ubiquitin carboxyl-terminal hydrolase isozyme L1 (UCHL1) rabbit polyclonal antibody (diluted 1:500; ab27053, Abcam), anti-glia maturation factor (GMFB) goat polyclonal antibody (diluted 1:200; sc46999, Santa Cruz Biotechnology), anti- / soluble N-ethylmaleimide-sensitive factor attachment protein ( , -SNAP) rabbit polyclonal antibody (diluted 1:1000; ab50483, Abcam), anti-17 -hydroxysteroid dehydrogenase type 10 (Hsd17b10) rabbit polyclonal antibody (diluted 1:500; ab17297, Abcam), anti- -enolase (ENO2) mouse monoclonal (85F11) antibody (diluted 1:2000; ab16808, Abcam), anti-peroxiredoxin 5 (PRDX5) rabbit polyclonal antibody (diluted 1:1500; ab16823, Abcam), anti-myelin basic protein (MBP) goat polyclonal antibody (diluted 1:200; sc13914, Santa Cruz Biotechnology), and anti-actin mouse monoclonal (clone C4) antibody (diluted 1:2500; MP Biomedicals).

    Techniques: Electrophoresis, Biomarker Discovery, Western Blot, Control, Comparison

    FIG. 5. Relationships between neurodegenerative disorders with the differentiated proteins in the biopsied brains compared with the control brains as predicted by PathwayStudio software. Proteins are shown as ovals (a dark gray oval means its altered tendency was consistent with the characteristic of neurodegenerative diseases; a light gray oval means its alteration may be compensation for neurode- generative disorders), and neurodegenerative disorders are represented by squares. Relationships are displayed with arrows and documented by literature citations (details listed in supplemental Table 2). CRYZ, -crystallin. FIG. 6. Immunohistochemistry analysis of ENO2, Hsd17b10, PRDX5, ,-SNAP, and GMFB in control and biopsied brains. ENO2, Hsd17b10, PRDX5, and ,-SNAP localized in the cytoplasm of neuron; GMFB localized in an astrocyte (black arrowheads indicate astrocyte in dentate gyrus). The trend in the differential expression of the proteins was in accordance with the variations obtained from 2DE and Western blotting.

    Journal: Molecular & Cellular Proteomics

    Article Title: Evaluation of Blastomere Biopsy Using a Mouse Model Indicates the Potential High Risk of Neurodegenerative Disorders in the Offspring

    doi: 10.1074/mcp.m800273-mcp200

    Figure Lengend Snippet: FIG. 5. Relationships between neurodegenerative disorders with the differentiated proteins in the biopsied brains compared with the control brains as predicted by PathwayStudio software. Proteins are shown as ovals (a dark gray oval means its altered tendency was consistent with the characteristic of neurodegenerative diseases; a light gray oval means its alteration may be compensation for neurode- generative disorders), and neurodegenerative disorders are represented by squares. Relationships are displayed with arrows and documented by literature citations (details listed in supplemental Table 2). CRYZ, -crystallin. FIG. 6. Immunohistochemistry analysis of ENO2, Hsd17b10, PRDX5, ,-SNAP, and GMFB in control and biopsied brains. ENO2, Hsd17b10, PRDX5, and ,-SNAP localized in the cytoplasm of neuron; GMFB localized in an astrocyte (black arrowheads indicate astrocyte in dentate gyrus). The trend in the differential expression of the proteins was in accordance with the variations obtained from 2DE and Western blotting.

    Article Snippet: Primary antibodies used were anti-ubiquitin carboxyl-terminal hydrolase isozyme L1 (UCHL1) rabbit polyclonal antibody (diluted 1:500; ab27053, Abcam), anti-glia maturation factor (GMFB) goat polyclonal antibody (diluted 1:200; sc46999, Santa Cruz Biotechnology), anti- / soluble N-ethylmaleimide-sensitive factor attachment protein ( , -SNAP) rabbit polyclonal antibody (diluted 1:1000; ab50483, Abcam), anti-17 -hydroxysteroid dehydrogenase type 10 (Hsd17b10) rabbit polyclonal antibody (diluted 1:500; ab17297, Abcam), anti- -enolase (ENO2) mouse monoclonal (85F11) antibody (diluted 1:2000; ab16808, Abcam), anti-peroxiredoxin 5 (PRDX5) rabbit polyclonal antibody (diluted 1:1500; ab16823, Abcam), anti-myelin basic protein (MBP) goat polyclonal antibody (diluted 1:200; sc13914, Santa Cruz Biotechnology), and anti-actin mouse monoclonal (clone C4) antibody (diluted 1:2500; MP Biomedicals).

    Techniques: Control, Software, Immunohistochemistry, Quantitative Proteomics, Western Blot