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Sino Biological biotin labeled her2 protein

ATPIF1 knockdown impairs in vitro antitumor activity and rewires the metabolism of CAR-T cells. (A) Schematic diagrams of lentiviral vector constructs for <t>Her2</t> CAR-T, Her2-IF1 CAR-T, and Her2-shIF1 CAR-T cells. a U6 promoter was inserted downstream of the Her2 CAR sequence to force the shRNA of ATPIF1. (B, C) ATPIF1 protein expression in CAR-T cells analyzed by Western blotting. (D) ATPIF1 mRNA levels in CAR-T cells determined by RT-qPCR. (E) Flow cytometric analysis of CAR-positive cells: CAR-T cells were first stained with biotin-labeled Her2 ectodomain protein, followed by incubation with PE-conjugated anti-biotin antibody for FACS detection of CAR + cells, the Y-axis was the count of cell numbers. (F) In vitro proliferation of CAR-T cells. The Her2 CAR-T, Her2-IF1 CAR-T and Her2-shIF1 CAR-T cells were cultivated with IL-2 stimulation (50 IU/ml) for 12 days, the numbers of CAR + cells were determined every two days. (G-I) Basal oxygen consumption rates (OCR) of resting Her2 CAR-T, Her2-IF1 CAR-T, and Her2-shIF1 CAR-T cells. Spare Respiratory Capacity (SRC) was calculated and plotted for both resting and activated cell states. (J, M) Cytolytic activity of untransfected T cells (UNT), Her2 CAR-T, Her2-IF1 CAR-T, and Her2-shIF1 CAR-T cells against SKBR3-Luc + cells at different effector-to-target (E:T) ratios following 24-hour co-culture under 21% O 2 and 1% O 2 conditions, respectively. (K, N) IL-2 concentration in co-culture supernatants from the experiment in (J, M) , respectively. (L, O) IFN-γ concentration in co-culture supernatants from the experiment in (J, M) , respectively. The ELISA method was used to determine the concentration of IL-2 and IFN-γ. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001..

Biotin Labeled Her2 Protein, supplied by Sino Biological, used in various techniques. Bioz Stars score: 95/100, based on 75 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/human+her2+erbb2+protein/Human+HER2+%2F+ErbB2+Protein/pmc12989527-107-19-24
Average 95 stars, based on 75 article reviews

biotin labeled her2 protein - by Bioz Stars, 2026-09

95/100 stars

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1) Product Images from "The critical role of Atpif1 in Her2-targeted CAR-T cell therapy for solid tumor via modulation of metabolism and mtDNA-STING signal pathway"

Article Title: The critical role of Atpif1 in Her2-targeted CAR-T cell therapy for solid tumor via modulation of metabolism and mtDNA-STING signal pathway

Journal: Frontiers in Immunology

doi: 10.3389/fimmu.2026.1733753

Figure Legend Snippet: ATPIF1 knockdown impairs in vitro antitumor activity and rewires the metabolism of CAR-T cells. (A) Schematic diagrams of lentiviral vector constructs for Her2 CAR-T, Her2-IF1 CAR-T, and Her2-shIF1 CAR-T cells. a U6 promoter was inserted downstream of the Her2 CAR sequence to force the shRNA of ATPIF1. (B, C) ATPIF1 protein expression in CAR-T cells analyzed by Western blotting. (D) ATPIF1 mRNA levels in CAR-T cells determined by RT-qPCR. (E) Flow cytometric analysis of CAR-positive cells: CAR-T cells were first stained with biotin-labeled Her2 ectodomain protein, followed by incubation with PE-conjugated anti-biotin antibody for FACS detection of CAR + cells, the Y-axis was the count of cell numbers. (F) In vitro proliferation of CAR-T cells. The Her2 CAR-T, Her2-IF1 CAR-T and Her2-shIF1 CAR-T cells were cultivated with IL-2 stimulation (50 IU/ml) for 12 days, the numbers of CAR + cells were determined every two days. (G-I) Basal oxygen consumption rates (OCR) of resting Her2 CAR-T, Her2-IF1 CAR-T, and Her2-shIF1 CAR-T cells. Spare Respiratory Capacity (SRC) was calculated and plotted for both resting and activated cell states. (J, M) Cytolytic activity of untransfected T cells (UNT), Her2 CAR-T, Her2-IF1 CAR-T, and Her2-shIF1 CAR-T cells against SKBR3-Luc + cells at different effector-to-target (E:T) ratios following 24-hour co-culture under 21% O 2 and 1% O 2 conditions, respectively. (K, N) IL-2 concentration in co-culture supernatants from the experiment in (J, M) , respectively. (L, O) IFN-γ concentration in co-culture supernatants from the experiment in (J, M) , respectively. The ELISA method was used to determine the concentration of IL-2 and IFN-γ. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001..

Techniques Used: Knockdown, In Vitro, Activity Assay, Plasmid Preparation, Construct, Sequencing, shRNA, Expressing, Western Blot, Quantitative RT-PCR, Staining, Labeling, Incubation, Co-Culture Assay, Concentration Assay, Enzyme-linked Immunosorbent Assay

ATPIF1 overexpression enhances the in vitro antitumor activity and oxygen consumption rate (OCR) of Her2-targeted CAR-T cells. (A) Schematic diagrams of lentiviral vector constructs for Her2 CAR-T and Her2-IF1 CAR-T cells. The ATPIF1 gene is integrated in tandem after the Her2 CAR vector. (B) Flow cytometric identification of GFP-positive cells. (C) Cytolytic activity of untransfected T cells (UNT), Her2 CAR-T, and Her2-IF1 CAR-T cells against SKBR3-Luc + cells at different effector-to-target (E:T) ratios following 24-hour co-culture. Lytic function was evaluated via bioluminescent killing assay in 96-well microplates. Data are presented as mean ± standard deviation (SD) of triplicate wells from three independent experiments. (D, E) Cytokines determination. Levels of IL-2 (D) and IFN-γ (E) in co-culture supernatants, measured by enzyme-linked immunosorbent assay (ELISA). (F–H) OXPHOS determination with the Seahorse equipment. Basal OCR of resting Her2 CAR-T and Her2-IF1 CAR-T cells. Spare Respiratory Capacity (SRC) was calculated and plotted for both resting and activated cell states. **P<0.01; ***P<0.001.

Figure Legend Snippet: ATPIF1 overexpression enhances the in vitro antitumor activity and oxygen consumption rate (OCR) of Her2-targeted CAR-T cells. (A) Schematic diagrams of lentiviral vector constructs for Her2 CAR-T and Her2-IF1 CAR-T cells. The ATPIF1 gene is integrated in tandem after the Her2 CAR vector. (B) Flow cytometric identification of GFP-positive cells. (C) Cytolytic activity of untransfected T cells (UNT), Her2 CAR-T, and Her2-IF1 CAR-T cells against SKBR3-Luc + cells at different effector-to-target (E:T) ratios following 24-hour co-culture. Lytic function was evaluated via bioluminescent killing assay in 96-well microplates. Data are presented as mean ± standard deviation (SD) of triplicate wells from three independent experiments. (D, E) Cytokines determination. Levels of IL-2 (D) and IFN-γ (E) in co-culture supernatants, measured by enzyme-linked immunosorbent assay (ELISA). (F–H) OXPHOS determination with the Seahorse equipment. Basal OCR of resting Her2 CAR-T and Her2-IF1 CAR-T cells. Spare Respiratory Capacity (SRC) was calculated and plotted for both resting and activated cell states. **P<0.01; ***P<0.001.

Techniques Used: Over Expression, In Vitro, Activity Assay, Plasmid Preparation, Construct, Co-Culture Assay, Standard Deviation, Enzyme-linked Immunosorbent Assay

ATPIF1 overexpression enhances CAR-T cell persistence but impairs in vivo antitumor activity. (A, B) Flow cytometric quantification of GFP + Her2 CAR-T and Her2-IF1 CAR-T cells in mouse peripheral blood mononuclear cells (PBMCs) and spleens. NCG mice (n=4 in each group) were intravenously injected with Her2 CAR-T, Her2-IF1 CAR-T cells (1x10 6 CAR-T cells per mouse). Three weeks later, the mice were euthanized, the peripheral blood and spleen were collected, respectively. The flow cytometry was used to detect the percentage of GFP + CAR-T cells. (C) Tumor growth inhibition by Her2 CAR-T and Her2-IF1 CAR-T cells in SKBR-3 xenografted NCG mice. NCG mice were xenografted with SKBR3 cells (n=5 or 6 in each group). When tumors reached ~100 mm³, mice were intravenously injected with untransfected T cells (UNT), Her2 CAR-T, Her2-IF1 CAR-T cells (1x10 6 CAR-T cells per mouse). Three weeks later, the mice were euthanized, tumors were excised and weighed. *P < 0.05; **P < 0.01. (D, F) The memory phenotype of Her2 CAR-T and Her2-IF1 CAR-T cells under 21% O 2 and 1% O 2 after stimulated with the conditional medium of SKBR-3 cells. (E, G) The exhaustion markers of PD-1, LAG-3 and TIM-3 determination in Her2 CAR-T and Her2-IF1 CAR-T cells under 21% O 2 and 1% O 2 after stimulated with the conditional medium of SKBR-3 cells. *P<0.05; **P<0.01; ***P<0.001.

Figure Legend Snippet: ATPIF1 overexpression enhances CAR-T cell persistence but impairs in vivo antitumor activity. (A, B) Flow cytometric quantification of GFP + Her2 CAR-T and Her2-IF1 CAR-T cells in mouse peripheral blood mononuclear cells (PBMCs) and spleens. NCG mice (n=4 in each group) were intravenously injected with Her2 CAR-T, Her2-IF1 CAR-T cells (1x10 6 CAR-T cells per mouse). Three weeks later, the mice were euthanized, the peripheral blood and spleen were collected, respectively. The flow cytometry was used to detect the percentage of GFP + CAR-T cells. (C) Tumor growth inhibition by Her2 CAR-T and Her2-IF1 CAR-T cells in SKBR-3 xenografted NCG mice. NCG mice were xenografted with SKBR3 cells (n=5 or 6 in each group). When tumors reached ~100 mm³, mice were intravenously injected with untransfected T cells (UNT), Her2 CAR-T, Her2-IF1 CAR-T cells (1x10 6 CAR-T cells per mouse). Three weeks later, the mice were euthanized, tumors were excised and weighed. *P < 0.05; **P < 0.01. (D, F) The memory phenotype of Her2 CAR-T and Her2-IF1 CAR-T cells under 21% O 2 and 1% O 2 after stimulated with the conditional medium of SKBR-3 cells. (E, G) The exhaustion markers of PD-1, LAG-3 and TIM-3 determination in Her2 CAR-T and Her2-IF1 CAR-T cells under 21% O 2 and 1% O 2 after stimulated with the conditional medium of SKBR-3 cells. *P<0.05; **P<0.01; ***P<0.001.

Techniques Used: Over Expression, In Vivo, Activity Assay, Injection, Flow Cytometry, Inhibition

Her2-shIF1 CAR-T cells exhibit enhanced in vivo antitumor activity. (A, B) In vivo tumor growth inhibition by UNT, Her2 CAR-T, Her2-IF1 CAR-T, and Her2-shIF1 CAR-T cells. NCG mice (n=5 or 6 in each group) were xenografted with SKBR3 cells. When tumors reached ~100 mm³, mice were intravenously injected with UNT, Her2 CAR-T, Her2-IF1 CAR-T cells (1x10 6 CAR-T cells per mouse). Two weeks later, the mice were euthanized, tumors were excised and weighed. (C) Tumor growth curves of xenografted tumors in each treatment group, the tumor size is calculated with the formula 0.5*a*b 2 , a is the length of tumor and b is the width of tumor. (D) Body weight changes of mice during the experimental period. (E) Spleen weights of mice at the end of the experiment. *P < 0.05; ***P < 0.001.

Figure Legend Snippet: Her2-shIF1 CAR-T cells exhibit enhanced in vivo antitumor activity. (A, B) In vivo tumor growth inhibition by UNT, Her2 CAR-T, Her2-IF1 CAR-T, and Her2-shIF1 CAR-T cells. NCG mice (n=5 or 6 in each group) were xenografted with SKBR3 cells. When tumors reached ~100 mm³, mice were intravenously injected with UNT, Her2 CAR-T, Her2-IF1 CAR-T cells (1x10 6 CAR-T cells per mouse). Two weeks later, the mice were euthanized, tumors were excised and weighed. (C) Tumor growth curves of xenografted tumors in each treatment group, the tumor size is calculated with the formula 0.5*a*b 2 , a is the length of tumor and b is the width of tumor. (D) Body weight changes of mice during the experimental period. (E) Spleen weights of mice at the end of the experiment. *P < 0.05; ***P < 0.001.

Techniques Used: In Vivo, Activity Assay, Inhibition, Injection

Immunofluorescence and multiplex immunohistochemistry analyses. (A, B) Enhanced CD3 + staining in group of Her2-shIF1 CAR-T in SKBR-3 xenografted tumor tissues. NCG mice were xenografted with SKBR3 cells (n=5 in each group); when tumors reached ~100 mm³, mice were intravenously injected with untransfected T cells (UNT), Her2 CAR-T, Her2-IF1 CAR-T, or Her2-shIF1 CAR-T cells. Four days later, Mice were euthanized, and tumors were excised for immunofluorescence staining. (C) Multiplex immunohistochemistry staining of 18 formalin-fixed paraffin-embedded (FFPE) human breast cancer tissue sections, showing ATPIF1 (yellow), CD3 (green), HIF-1α (orange), and DAPI (blue). (D, E) Quantitative analysis of the correlation between CD3 + staining and ATPIF1 + cells (D) , and between HIF-1α + and ATPIF1 + cells (E) from the results of Multiplex immunohistochemistry staining in (C) respectively. **P<0.01, ***P<0.001.

Figure Legend Snippet: Immunofluorescence and multiplex immunohistochemistry analyses. (A, B) Enhanced CD3 + staining in group of Her2-shIF1 CAR-T in SKBR-3 xenografted tumor tissues. NCG mice were xenografted with SKBR3 cells (n=5 in each group); when tumors reached ~100 mm³, mice were intravenously injected with untransfected T cells (UNT), Her2 CAR-T, Her2-IF1 CAR-T, or Her2-shIF1 CAR-T cells. Four days later, Mice were euthanized, and tumors were excised for immunofluorescence staining. (C) Multiplex immunohistochemistry staining of 18 formalin-fixed paraffin-embedded (FFPE) human breast cancer tissue sections, showing ATPIF1 (yellow), CD3 (green), HIF-1α (orange), and DAPI (blue). (D, E) Quantitative analysis of the correlation between CD3 + staining and ATPIF1 + cells (D) , and between HIF-1α + and ATPIF1 + cells (E) from the results of Multiplex immunohistochemistry staining in (C) respectively. **P<0.01, ***P<0.001.

Techniques Used: Immunofluorescence, Multiplex Assay, Immunohistochemistry, Staining, Injection, Formalin-fixed Paraffin-Embedded

ATPIF1 knockdown induces mPTP opening, mtDNA leakage, and STING signaling pathway activation. (A, B) Flow cytometric detection of mPTP opening using Calcein-AM staining. The Her2 CAR-T, Her2-IF1 CAR-T and Her-shIF1 CAR-T cells were treated with or without N-Acetylcysteine (NAC, 10 mM) for 24 hours, and then stained with Calcein-AM, flow cytometry was used to detect the content of Calcein-AM, which indicated the levels of mPTP opening. (C) Western blot analysis of p-STING, total STING, and VDAC protein expression. (D) Quantification of p-STING relative to total STING. (E) Quantification of VDAC relative to β-actin (loading control). (F–H) RT-qPCR analysis of cytoplasmic mtDNA (ND-1 and D-loop) and mRNA levels of IFI44 and IFN-β. Cytoplasmic RNA was extracted for detection. (I) Generation of ρ 0 CAR-T cells via ethidium bromide (EB) treatment for 2 weeks. Total mtDNA was extracted to detect D-loop levels. (J) RT-qPCR analysis of IFI44 mRNA expression. *P < 0.05; **P < 0.01; ***P < 0.001; ****P<0.0001.

Figure Legend Snippet: ATPIF1 knockdown induces mPTP opening, mtDNA leakage, and STING signaling pathway activation. (A, B) Flow cytometric detection of mPTP opening using Calcein-AM staining. The Her2 CAR-T, Her2-IF1 CAR-T and Her-shIF1 CAR-T cells were treated with or without N-Acetylcysteine (NAC, 10 mM) for 24 hours, and then stained with Calcein-AM, flow cytometry was used to detect the content of Calcein-AM, which indicated the levels of mPTP opening. (C) Western blot analysis of p-STING, total STING, and VDAC protein expression. (D) Quantification of p-STING relative to total STING. (E) Quantification of VDAC relative to β-actin (loading control). (F–H) RT-qPCR analysis of cytoplasmic mtDNA (ND-1 and D-loop) and mRNA levels of IFI44 and IFN-β. Cytoplasmic RNA was extracted for detection. (I) Generation of ρ 0 CAR-T cells via ethidium bromide (EB) treatment for 2 weeks. Total mtDNA was extracted to detect D-loop levels. (J) RT-qPCR analysis of IFI44 mRNA expression. *P < 0.05; **P < 0.01; ***P < 0.001; ****P<0.0001.

Techniques Used: Knockdown, Activation Assay, Staining, Flow Cytometry, Western Blot, Expressing, Control, Quantitative RT-PCR

The enhanced in vivo antitumor activity of Her2-shIF1 CAR-T cells is dependent on STING activation. (A, B) Transwell migration assay of CAR-T cells under hypoxic conditions (1% O 2 ) with or without H151 inhibition. CAR-T cells were stained with Cell-Tracker Red CMTPX and seeded into the upper Transwell chamber. Migrated cells in the lower chamber were visualized via fluorescence microscopy, with five random fields per well imaged for quantification. (C, D) Excised tumor size and weight following treatment with different CAR-T cells, either alone or in combination with H151. NCG mice were xenografted with SKBR3 cells (n=5 in each group). When tumors reached ~100 mm³, mice were intravenously injected with untransfected T cells (UNT), Her2 CAR-T, Her2-IF1 CAR-T, or Her2-shIF1 CAR-T cells (1x10 6 CAR-T cells per mouse); or treated with or without H151 via intraperitoneal injection (7 mg/kg) as indicated. Two weeks later, the mice were euthanized, tumors were excised and weighed. **P < 0.01; ***P < 0.001.

Figure Legend Snippet: The enhanced in vivo antitumor activity of Her2-shIF1 CAR-T cells is dependent on STING activation. (A, B) Transwell migration assay of CAR-T cells under hypoxic conditions (1% O 2 ) with or without H151 inhibition. CAR-T cells were stained with Cell-Tracker Red CMTPX and seeded into the upper Transwell chamber. Migrated cells in the lower chamber were visualized via fluorescence microscopy, with five random fields per well imaged for quantification. (C, D) Excised tumor size and weight following treatment with different CAR-T cells, either alone or in combination with H151. NCG mice were xenografted with SKBR3 cells (n=5 in each group). When tumors reached ~100 mm³, mice were intravenously injected with untransfected T cells (UNT), Her2 CAR-T, Her2-IF1 CAR-T, or Her2-shIF1 CAR-T cells (1x10 6 CAR-T cells per mouse); or treated with or without H151 via intraperitoneal injection (7 mg/kg) as indicated. Two weeks later, the mice were euthanized, tumors were excised and weighed. **P < 0.01; ***P < 0.001.

Techniques Used: In Vivo, Activity Assay, Activation Assay, Transwell Migration Assay, Inhibition, Staining, Fluorescence, Microscopy, Injection

Expressing:

Article Title: Chimeric Antigen Cytotoxic Receptors for In-Vivo Engineering of Tumor-targeting Natural Killer Cells
Article Snippet: .. A human HER2/ERBB2 protein-His tag (10004-H08H-100, Sino Biological) conjugated with Alexa Fluor 647 was used to determine the expression of HER2-CAR in transfected cells, and viability was determined by 7-AAD labeling. ..

Article Title: Chimeric Antigen Cytotoxic Receptors for In Vivo Engineering of Tumor-Targeting NK Cells.
Article Snippet: .. A human HER2/ERBB2 protein-His tag (Sino Biological) conjugated with Alexa Fluor 647 (1:200 dilution) was used to determine the expression of HER2-CAR in transfected cells, and viability was determined by 7-aminoactinomycin D (7-AAD) labeling (1:1000 dilution). ..

Article Title: CAR-macrophage therapy for HER2-overexpressing advanced solid tumors: a phase 1 trial.
Article Snippet: Chimeric antigen receptor (CAR) macrophages (CAR-Ms) mediate antitumor immunity via phagocytosis, cytokine release, activation of the tumor microenvironment and antigen presentation.. We report results from a non-prespecified interim analysis of a first-in-human, phase 1 clinical trial of CT-0508, an anti-human epidermal growth factor receptor 2 (HER2) CAR-M in patients with advanced HER2-overexpressing tumors.. Fourteen patients were treated across two different regimens.

Article Title: Chimeric Antigen Cytotoxic Receptors for In Vivo Engineering of Tumor-Targeting NK Cells
Article Snippet: .. A human HER2/ERBB2 protein-His tag (Sino Biological) conjugated with Alexa Fluor 647 (1:200 dilution) was used to determine the expression of HER2-CAR in transfected cells, and viability was determined by 7-aminoactinomycin D (7-AAD) labeling (1:1000 dilution). ..

Article Title: Enhanced anti-tumor effects through continuous administration of engineered CAR-macrophages derived from pluripotent stem cell-derived myeloid cell lines
Article Snippet: The following antibodies were used in this study flow cytometry analysis: CD36-FITC (#IM0766U; Beckman Coulter, Brea, CA, USA), CD45-FITC (#368508; BioLegend, San Diego, CA, USA), HLA-DR-FITC (#307604; Bio Legend), CD206-FITC (#551135; BD bioscience), CD86-FITC (#555657; BD bioscience), CD11b-Alexa488 (#557701; BD bioscience), CD340(erbB2/HER-2)-PE (#324406; Bio Legend), CD14-APC (#IM2580; Beckman Coulter), CD11c-APC (559877; BD bioscience), Fc Receptor Blocking Solution (#422302; Bio Legend), FITC IgG1 isotype control (#400110; Bio Legend), FITC IgG2a isotype control (#349051; BD bioscience), PE IgG1 isotype control (#400112; Bio Legend), APC IgG1 isotype control (#400122; Bio Legend), and APC IgG2a isotype control (#400512; Bio Legend). .. For confirmation of anti-HER2 CAR expression, primary staining was performed with human HER2/ERBB2 Protein-His tag (#10004H08H100; Sino Biological, Beijing, China), followed by secondary staining with anti-His Tag APC (#IC050A; R&D Systems). ..

Transfection:

Labeling:

other:

Article Title: Paper discs in a 3D printed microplate hybrid microfluidic device for low-cost, rapid, and ultrasensitive paper-based bioluminescence detection of human epidermal growth factor receptor 2 (HER2) breast cancer biomarker
Article Snippet: Human HER2/ErbB2 protein (His Tag) (catalog number: 10004-H08H), HER2/ErbB2 antibody - rabbit Mab (catalog number: 10004-R205) as a detection antibody, and HER2/ErbB2/CD340 antibody - rabbit Mab (catalog number: 10004-R511) as capture antibody were purchased from Sino Biological (USA).

Staining:

Article Title: Activation of FCGR2A enhances the antitumor efficacy of hPSC-derived CAR-M
Article Snippet: .. Cells were first incubated with Human HER2/ERBB2 Protein His tag (Sino Biological) as the primary stain, followed by Human Anti-His Tag APC (R&D Systems) as the secondary stain. ..

Incubation:

Recombinant:

Article Title: Synergistic Role of MoO₃–CTAB Nanostructures in Boosting Electrochemical Immunosensing of HER2
Article Snippet: Cancer is a disease that impacts a significant portion of the global population, and delayed diagnosis complicates treatment and elevates mortality rates.. The disease affects many organs and has many subtypes.. Breast cancer is the most prevalent condition among women, and biomarkers are crucial for disease diagnosis.

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