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CD8 + T cells control tumor growth following targeting the RGDKGE collagen peptide. C57BL/6 mice were treated intraperitoneally with function-blocking <t>anti–CD8α-depleting</t> antibody <t>(BP0061)</t> or non-specific control antibodies. A: Example of flow cytometry analysis for CD8 + T cells from spleens of mice treated with non-specific control antibody (Ab Cont) or anti–CD8α-depleting antibody (Anti-CD8). B: Quantification of CD8 + T cells from spleens of mice treated with non-specific control antibody (Ab Cont) or anti–CD8α-depleting antibody (Anti-CD8). Data represent CD8 + T cells from four mice per group. C: Example of flow cytometry analysis for CD4 + T cells from spleens of mice treated with non-specific control antibody (Ab Cont) or anti–CD8α-depleting antibody (Anti-CD8). D: Quantification of CD4 + T cells from spleens of mice treated with either non-specific control antibody (Ab Cont) or anti–CD8α-depleting antibody (Anti-CD8). Data represent CD4 + T cells from four mice per group. E: Examples of CD8 + T cells (red) in B16F10 tumor section from either non-specific control antibody (Ab Cont) or anti–CD8α-depleting antibody (Anti-CD8) treated mice. F: Quantification of the mean CD8 + T-cell count from four different tumors from each treatment group using five to seven ×200 microscopic fields from each tumor. Data represent CD8 + T-cell counts from four different tumors from each treatment group. G: Quantification of B16F10 tumor size from control antibody-depleted (Ab Cont) or CD8-depleted (Anti-CD8) mice over 14 days. Data points represent tumor volume from four mice per condition. H: Quantification of B16F10 tumor size from control antibody-depleted mice treated with non-specific control antibody (Ab Cont) or anti-RGDKGE antibody [monoclonal antibody (Mab) XL313] over 14 days. Data points represent tumor volume from eight mice per condition. I: Quantification of B16F10 tumor size from CD8-depleted mice treated with non-specific control antibody (Ab Cont) or anti-RGDKGE antibody (Mab XL313) over 14 days. Data points represent tumor volume from seven to eight mice per condition. Data are given as means ± SEM ( B , D , and F – I ). ∗ P < 0.05, ∗∗∗ P < 0.001, and ∗∗∗∗ P < 0.0001. Scale bars = 50 μm ( E ). SSC, side scatter.
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CD8 + T cells control tumor growth following targeting the RGDKGE collagen peptide. C57BL/6 mice were treated intraperitoneally with function-blocking <t>anti–CD8α-depleting</t> antibody <t>(BP0061)</t> or non-specific control antibodies. A: Example of flow cytometry analysis for CD8 + T cells from spleens of mice treated with non-specific control antibody (Ab Cont) or anti–CD8α-depleting antibody (Anti-CD8). B: Quantification of CD8 + T cells from spleens of mice treated with non-specific control antibody (Ab Cont) or anti–CD8α-depleting antibody (Anti-CD8). Data represent CD8 + T cells from four mice per group. C: Example of flow cytometry analysis for CD4 + T cells from spleens of mice treated with non-specific control antibody (Ab Cont) or anti–CD8α-depleting antibody (Anti-CD8). D: Quantification of CD4 + T cells from spleens of mice treated with either non-specific control antibody (Ab Cont) or anti–CD8α-depleting antibody (Anti-CD8). Data represent CD4 + T cells from four mice per group. E: Examples of CD8 + T cells (red) in B16F10 tumor section from either non-specific control antibody (Ab Cont) or anti–CD8α-depleting antibody (Anti-CD8) treated mice. F: Quantification of the mean CD8 + T-cell count from four different tumors from each treatment group using five to seven ×200 microscopic fields from each tumor. Data represent CD8 + T-cell counts from four different tumors from each treatment group. G: Quantification of B16F10 tumor size from control antibody-depleted (Ab Cont) or CD8-depleted (Anti-CD8) mice over 14 days. Data points represent tumor volume from four mice per condition. H: Quantification of B16F10 tumor size from control antibody-depleted mice treated with non-specific control antibody (Ab Cont) or anti-RGDKGE antibody [monoclonal antibody (Mab) XL313] over 14 days. Data points represent tumor volume from eight mice per condition. I: Quantification of B16F10 tumor size from CD8-depleted mice treated with non-specific control antibody (Ab Cont) or anti-RGDKGE antibody (Mab XL313) over 14 days. Data points represent tumor volume from seven to eight mice per condition. Data are given as means ± SEM ( B , D , and F – I ). ∗ P < 0.05, ∗∗∗ P < 0.001, and ∗∗∗∗ P < 0.0001. Scale bars = 50 μm ( E ). SSC, side scatter.
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CD8 + T cells control tumor growth following targeting the RGDKGE collagen peptide. C57BL/6 mice were treated intraperitoneally with function-blocking <t>anti–CD8α-depleting</t> antibody <t>(BP0061)</t> or non-specific control antibodies. A: Example of flow cytometry analysis for CD8 + T cells from spleens of mice treated with non-specific control antibody (Ab Cont) or anti–CD8α-depleting antibody (Anti-CD8). B: Quantification of CD8 + T cells from spleens of mice treated with non-specific control antibody (Ab Cont) or anti–CD8α-depleting antibody (Anti-CD8). Data represent CD8 + T cells from four mice per group. C: Example of flow cytometry analysis for CD4 + T cells from spleens of mice treated with non-specific control antibody (Ab Cont) or anti–CD8α-depleting antibody (Anti-CD8). D: Quantification of CD4 + T cells from spleens of mice treated with either non-specific control antibody (Ab Cont) or anti–CD8α-depleting antibody (Anti-CD8). Data represent CD4 + T cells from four mice per group. E: Examples of CD8 + T cells (red) in B16F10 tumor section from either non-specific control antibody (Ab Cont) or anti–CD8α-depleting antibody (Anti-CD8) treated mice. F: Quantification of the mean CD8 + T-cell count from four different tumors from each treatment group using five to seven ×200 microscopic fields from each tumor. Data represent CD8 + T-cell counts from four different tumors from each treatment group. G: Quantification of B16F10 tumor size from control antibody-depleted (Ab Cont) or CD8-depleted (Anti-CD8) mice over 14 days. Data points represent tumor volume from four mice per condition. H: Quantification of B16F10 tumor size from control antibody-depleted mice treated with non-specific control antibody (Ab Cont) or anti-RGDKGE antibody [monoclonal antibody (Mab) XL313] over 14 days. Data points represent tumor volume from eight mice per condition. I: Quantification of B16F10 tumor size from CD8-depleted mice treated with non-specific control antibody (Ab Cont) or anti-RGDKGE antibody (Mab XL313) over 14 days. Data points represent tumor volume from seven to eight mice per condition. Data are given as means ± SEM ( B , D , and F – I ). ∗ P < 0.05, ∗∗∗ P < 0.001, and ∗∗∗∗ P < 0.0001. Scale bars = 50 μm ( E ). SSC, side scatter.
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CD8 + T cells control tumor growth following targeting the RGDKGE collagen peptide. C57BL/6 mice were treated intraperitoneally with function-blocking <t>anti–CD8α-depleting</t> antibody <t>(BP0061)</t> or non-specific control antibodies. A: Example of flow cytometry analysis for CD8 + T cells from spleens of mice treated with non-specific control antibody (Ab Cont) or anti–CD8α-depleting antibody (Anti-CD8). B: Quantification of CD8 + T cells from spleens of mice treated with non-specific control antibody (Ab Cont) or anti–CD8α-depleting antibody (Anti-CD8). Data represent CD8 + T cells from four mice per group. C: Example of flow cytometry analysis for CD4 + T cells from spleens of mice treated with non-specific control antibody (Ab Cont) or anti–CD8α-depleting antibody (Anti-CD8). D: Quantification of CD4 + T cells from spleens of mice treated with either non-specific control antibody (Ab Cont) or anti–CD8α-depleting antibody (Anti-CD8). Data represent CD4 + T cells from four mice per group. E: Examples of CD8 + T cells (red) in B16F10 tumor section from either non-specific control antibody (Ab Cont) or anti–CD8α-depleting antibody (Anti-CD8) treated mice. F: Quantification of the mean CD8 + T-cell count from four different tumors from each treatment group using five to seven ×200 microscopic fields from each tumor. Data represent CD8 + T-cell counts from four different tumors from each treatment group. G: Quantification of B16F10 tumor size from control antibody-depleted (Ab Cont) or CD8-depleted (Anti-CD8) mice over 14 days. Data points represent tumor volume from four mice per condition. H: Quantification of B16F10 tumor size from control antibody-depleted mice treated with non-specific control antibody (Ab Cont) or anti-RGDKGE antibody [monoclonal antibody (Mab) XL313] over 14 days. Data points represent tumor volume from eight mice per condition. I: Quantification of B16F10 tumor size from CD8-depleted mice treated with non-specific control antibody (Ab Cont) or anti-RGDKGE antibody (Mab XL313) over 14 days. Data points represent tumor volume from seven to eight mice per condition. Data are given as means ± SEM ( B , D , and F – I ). ∗ P < 0.05, ∗∗∗ P < 0.001, and ∗∗∗∗ P < 0.0001. Scale bars = 50 μm ( E ). SSC, side scatter.
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CD8 + T cells control tumor growth following targeting the RGDKGE collagen peptide. C57BL/6 mice were treated intraperitoneally with function-blocking <t>anti–CD8α-depleting</t> antibody <t>(BP0061)</t> or non-specific control antibodies. A: Example of flow cytometry analysis for CD8 + T cells from spleens of mice treated with non-specific control antibody (Ab Cont) or anti–CD8α-depleting antibody (Anti-CD8). B: Quantification of CD8 + T cells from spleens of mice treated with non-specific control antibody (Ab Cont) or anti–CD8α-depleting antibody (Anti-CD8). Data represent CD8 + T cells from four mice per group. C: Example of flow cytometry analysis for CD4 + T cells from spleens of mice treated with non-specific control antibody (Ab Cont) or anti–CD8α-depleting antibody (Anti-CD8). D: Quantification of CD4 + T cells from spleens of mice treated with either non-specific control antibody (Ab Cont) or anti–CD8α-depleting antibody (Anti-CD8). Data represent CD4 + T cells from four mice per group. E: Examples of CD8 + T cells (red) in B16F10 tumor section from either non-specific control antibody (Ab Cont) or anti–CD8α-depleting antibody (Anti-CD8) treated mice. F: Quantification of the mean CD8 + T-cell count from four different tumors from each treatment group using five to seven ×200 microscopic fields from each tumor. Data represent CD8 + T-cell counts from four different tumors from each treatment group. G: Quantification of B16F10 tumor size from control antibody-depleted (Ab Cont) or CD8-depleted (Anti-CD8) mice over 14 days. Data points represent tumor volume from four mice per condition. H: Quantification of B16F10 tumor size from control antibody-depleted mice treated with non-specific control antibody (Ab Cont) or anti-RGDKGE antibody [monoclonal antibody (Mab) XL313] over 14 days. Data points represent tumor volume from eight mice per condition. I: Quantification of B16F10 tumor size from CD8-depleted mice treated with non-specific control antibody (Ab Cont) or anti-RGDKGE antibody (Mab XL313) over 14 days. Data points represent tumor volume from seven to eight mice per condition. Data are given as means ± SEM ( B , D , and F – I ). ∗ P < 0.05, ∗∗∗ P < 0.001, and ∗∗∗∗ P < 0.0001. Scale bars = 50 μm ( E ). SSC, side scatter.
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CD8 + T cells control tumor growth following targeting the RGDKGE collagen peptide. C57BL/6 mice were treated intraperitoneally with function-blocking <t>anti–CD8α-depleting</t> antibody <t>(BP0061)</t> or non-specific control antibodies. A: Example of flow cytometry analysis for CD8 + T cells from spleens of mice treated with non-specific control antibody (Ab Cont) or anti–CD8α-depleting antibody (Anti-CD8). B: Quantification of CD8 + T cells from spleens of mice treated with non-specific control antibody (Ab Cont) or anti–CD8α-depleting antibody (Anti-CD8). Data represent CD8 + T cells from four mice per group. C: Example of flow cytometry analysis for CD4 + T cells from spleens of mice treated with non-specific control antibody (Ab Cont) or anti–CD8α-depleting antibody (Anti-CD8). D: Quantification of CD4 + T cells from spleens of mice treated with either non-specific control antibody (Ab Cont) or anti–CD8α-depleting antibody (Anti-CD8). Data represent CD4 + T cells from four mice per group. E: Examples of CD8 + T cells (red) in B16F10 tumor section from either non-specific control antibody (Ab Cont) or anti–CD8α-depleting antibody (Anti-CD8) treated mice. F: Quantification of the mean CD8 + T-cell count from four different tumors from each treatment group using five to seven ×200 microscopic fields from each tumor. Data represent CD8 + T-cell counts from four different tumors from each treatment group. G: Quantification of B16F10 tumor size from control antibody-depleted (Ab Cont) or CD8-depleted (Anti-CD8) mice over 14 days. Data points represent tumor volume from four mice per condition. H: Quantification of B16F10 tumor size from control antibody-depleted mice treated with non-specific control antibody (Ab Cont) or anti-RGDKGE antibody [monoclonal antibody (Mab) XL313] over 14 days. Data points represent tumor volume from eight mice per condition. I: Quantification of B16F10 tumor size from CD8-depleted mice treated with non-specific control antibody (Ab Cont) or anti-RGDKGE antibody (Mab XL313) over 14 days. Data points represent tumor volume from seven to eight mice per condition. Data are given as means ± SEM ( B , D , and F – I ). ∗ P < 0.05, ∗∗∗ P < 0.001, and ∗∗∗∗ P < 0.0001. Scale bars = 50 μm ( E ). SSC, side scatter.
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CD8 + T cells control tumor growth following targeting the RGDKGE collagen peptide. C57BL/6 mice were treated intraperitoneally with function-blocking anti–CD8α-depleting antibody (BP0061) or non-specific control antibodies. A: Example of flow cytometry analysis for CD8 + T cells from spleens of mice treated with non-specific control antibody (Ab Cont) or anti–CD8α-depleting antibody (Anti-CD8). B: Quantification of CD8 + T cells from spleens of mice treated with non-specific control antibody (Ab Cont) or anti–CD8α-depleting antibody (Anti-CD8). Data represent CD8 + T cells from four mice per group. C: Example of flow cytometry analysis for CD4 + T cells from spleens of mice treated with non-specific control antibody (Ab Cont) or anti–CD8α-depleting antibody (Anti-CD8). D: Quantification of CD4 + T cells from spleens of mice treated with either non-specific control antibody (Ab Cont) or anti–CD8α-depleting antibody (Anti-CD8). Data represent CD4 + T cells from four mice per group. E: Examples of CD8 + T cells (red) in B16F10 tumor section from either non-specific control antibody (Ab Cont) or anti–CD8α-depleting antibody (Anti-CD8) treated mice. F: Quantification of the mean CD8 + T-cell count from four different tumors from each treatment group using five to seven ×200 microscopic fields from each tumor. Data represent CD8 + T-cell counts from four different tumors from each treatment group. G: Quantification of B16F10 tumor size from control antibody-depleted (Ab Cont) or CD8-depleted (Anti-CD8) mice over 14 days. Data points represent tumor volume from four mice per condition. H: Quantification of B16F10 tumor size from control antibody-depleted mice treated with non-specific control antibody (Ab Cont) or anti-RGDKGE antibody [monoclonal antibody (Mab) XL313] over 14 days. Data points represent tumor volume from eight mice per condition. I: Quantification of B16F10 tumor size from CD8-depleted mice treated with non-specific control antibody (Ab Cont) or anti-RGDKGE antibody (Mab XL313) over 14 days. Data points represent tumor volume from seven to eight mice per condition. Data are given as means ± SEM ( B , D , and F – I ). ∗ P < 0.05, ∗∗∗ P < 0.001, and ∗∗∗∗ P < 0.0001. Scale bars = 50 μm ( E ). SSC, side scatter.

Journal: The American Journal of Pathology

Article Title: Inhibiting the Secreted RGDKGE Collagen Peptide Selectively Controls CD8 + T-Cell Migration on Denatured Collagen-IV and Enhances Their Accumulation in Tumors

doi: 10.1016/j.ajpath.2025.09.008

Figure Lengend Snippet: CD8 + T cells control tumor growth following targeting the RGDKGE collagen peptide. C57BL/6 mice were treated intraperitoneally with function-blocking anti–CD8α-depleting antibody (BP0061) or non-specific control antibodies. A: Example of flow cytometry analysis for CD8 + T cells from spleens of mice treated with non-specific control antibody (Ab Cont) or anti–CD8α-depleting antibody (Anti-CD8). B: Quantification of CD8 + T cells from spleens of mice treated with non-specific control antibody (Ab Cont) or anti–CD8α-depleting antibody (Anti-CD8). Data represent CD8 + T cells from four mice per group. C: Example of flow cytometry analysis for CD4 + T cells from spleens of mice treated with non-specific control antibody (Ab Cont) or anti–CD8α-depleting antibody (Anti-CD8). D: Quantification of CD4 + T cells from spleens of mice treated with either non-specific control antibody (Ab Cont) or anti–CD8α-depleting antibody (Anti-CD8). Data represent CD4 + T cells from four mice per group. E: Examples of CD8 + T cells (red) in B16F10 tumor section from either non-specific control antibody (Ab Cont) or anti–CD8α-depleting antibody (Anti-CD8) treated mice. F: Quantification of the mean CD8 + T-cell count from four different tumors from each treatment group using five to seven ×200 microscopic fields from each tumor. Data represent CD8 + T-cell counts from four different tumors from each treatment group. G: Quantification of B16F10 tumor size from control antibody-depleted (Ab Cont) or CD8-depleted (Anti-CD8) mice over 14 days. Data points represent tumor volume from four mice per condition. H: Quantification of B16F10 tumor size from control antibody-depleted mice treated with non-specific control antibody (Ab Cont) or anti-RGDKGE antibody [monoclonal antibody (Mab) XL313] over 14 days. Data points represent tumor volume from eight mice per condition. I: Quantification of B16F10 tumor size from CD8-depleted mice treated with non-specific control antibody (Ab Cont) or anti-RGDKGE antibody (Mab XL313) over 14 days. Data points represent tumor volume from seven to eight mice per condition. Data are given as means ± SEM ( B , D , and F – I ). ∗ P < 0.05, ∗∗∗ P < 0.001, and ∗∗∗∗ P < 0.0001. Scale bars = 50 μm ( E ). SSC, side scatter.

Article Snippet: Mab XL313, functional blocking anti-CD8α (BP0061) used for depletion, and non-specific control antibodies were acquired from Bio X-Cell (Lebanon, NH).

Techniques: Control, Blocking Assay, Flow Cytometry, Cell Characterization