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Anti cd8α clone 53 6.72

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Anti-CD8α (clone 53–6.72) is a monoclonal antibody that recognizes the CD8α chain, a component of the CD8 co-receptor expressed on the surface of cytotoxic T cells and a subset of natural killer cells. This antibody can be used for the identification and isolation of CD8+ T cells in various applications.

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Lab products found in correlation

Anti cd8β clone 53 5.8, by BioXCell (2 mentions) Rat igg2a clone 2a3, by BioXCell (2 mentions) Cd122, by BioLegend (1 mentions) Klrg1, by BioLegend (1 mentions) Goat anti rat igg, by Jackson ImmunoResearch (1 mentions) Moma 1, by Bio-Rad (1 mentions) Mhcii, by BioLegend (1 mentions) C57bl 6 mice, by Jackson ImmunoResearch (1 mentions) Cd62l, by BioLegend (1 mentions) Anti cd4 clone gk1, by BioXCell (1 mentions)

Spelling variants of this product

Clone 53–6.7 (10 citations) Anti-CD8α (9 citations)

5 protocols using anti cd8α clone 53 6.72

1

OT1 TCR Transgenic Mouse Protocol

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All experiments were performed with 5–10 week old C57BL/6 mice of either sex, purchased from Jackson Laboratories. OT1 are a TCR transgenic mouse specific for the SIINFEKL peptide derived from ovalbumin (amino acids 257–254) in the context of H-2Kb and were bred and housed at the University of Colorado vivarium. All animal protocols were approved by the Institute of Animal Care and Use Committees of the University of Colorado. Antibodies used for depletion, anti-CD8α (clone 53–6.72) and anti-CD8β (clone 53–5.8), were purchased from Bio X Cell and/or made in house. Fluorochrome-conjugated antibodies used for flow cytometry include CD8α, CD8β, B220, CD44, CD62L, MHC-II, CD45.1, CD45.2, CD122, CD127, Eomesodermin and KLRG1 all purchased from Biolegend and Goat anti-Rat IgG from Jackson Immunoresearch Laboratories. Fluorochrome-conjugated antibodies used for microscopy include MOMA-1 (BioRad), IgM (in house), and CD3ε and CD45.1 (Biolegend). SIINFEKL and SIYNFEKL peptide used for in vitro stimulation or in vivo vaccination was synthesized by the University of Colorado Protein Production Shared Resource facility.
Cross E.W., Blain T.J., Mathew D, & Kedl R.M. (2019). Anti-CD8 monoclonal antibody-mediated depletion alters the phenotype and behavior of surviving CD8+ T cells. PLoS ONE, 14(2), e0211446.
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2

Syngeneic Murine Tumor Implantation

Cited 2 times
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Cells were injected intradermally onto the backs of C57BL/6, NOD.Cg-PrkdcscidIl2rgtm1Wjl/SzJ (NSG), or B6. 129S7-Rag1tm1Mom/J (Rag1 KO) mice. Cell numbers were based on previous publications and past experiments (YUMM1.1 = 1×106 cells; 1014 = 5×105 cells) (Meeth et al., 2016 (link)). Tumors were considered fully formed when they reached ~50mm3. For CD8+ depletion, animals were treated with 300 μg of anti-CD8α (clone 53–6.72) or the corresponding isotype control (Rat IgG2a, clone 2A3) (BioXCell; West Lebanon, NH) by intraperitoneal injection 2 days prior to tumor implantation and 2 times per week for the duration of the experiment. Treatments were determined based on previous publications. Animals were sacrificed when tumors exceeded 650mm3 (Erkes et al., 2020 (link)).
Rosenbaum S.R., Tiago M., Caksa S., Capparelli C., Purwin T.J., Kumar G., Glasheen M., Pomante D., Kotas D., Chervoneva I, & Aplin A.E. (2021). SOX10 requirement for melanoma tumor growth is due, in part, to immune-mediated effects. Cell reports, 37(10), 110085.
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3

Syngeneic Murine Tumor Implantation

Cited 2 times
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Cells were injected intradermally onto the backs of C57BL/6, NOD.Cg-PrkdcscidIl2rgtm1Wjl/SzJ (NSG), or B6. 129S7-Rag1tm1Mom/J (Rag1 KO) mice. Cell numbers were based on previous publications and past experiments (YUMM1.1 = 1×106 cells; 1014 = 5×105 cells) (Meeth et al., 2016 (link)). Tumors were considered fully formed when they reached ~50mm3. For CD8+ depletion, animals were treated with 300 μg of anti-CD8α (clone 53–6.72) or the corresponding isotype control (Rat IgG2a, clone 2A3) (BioXCell; West Lebanon, NH) by intraperitoneal injection 2 days prior to tumor implantation and 2 times per week for the duration of the experiment. Treatments were determined based on previous publications. Animals were sacrificed when tumors exceeded 650mm3 (Erkes et al., 2020 (link)).
Rosenbaum S.R., Tiago M., Caksa S., Capparelli C., Purwin T.J., Kumar G., Glasheen M., Pomante D., Kotas D., Chervoneva I, & Aplin A.E. (2021). SOX10 requirement for melanoma tumor growth is due, in part, to immune-mediated effects. Cell reports, 37(10), 110085.
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4

T-cell Depletion and Leukemia Progression

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T‐cells were depleted from aged (24 months) C57BL/6 mice using CD4 (anti‐CD4; clone GK1.5; purchased from Bio X Cell) and CD8 (anti‐CD8α; clone 53–6.72 and anti‐CD8β; clone 53–5.8; purchased from Bio X Cell) T‐cell depleting antibodies (i.p.; 0.5 mg/mouse/each antibody for two consecutive days). One day after the administration of T‐cell depleting antibodies, mice were treated with Control Ig (i.v.; 100 µg/mouse) or rIL‐37 (i.v.; 100 µg/mouse), and these treatments continued once weekly for the duration of the experiment. Two days after the administration of T‐cell depleting antibodies, mice were transplanted with murine B‐ALL (GFP‐expressing, BCR‐ABL1+/Arfnull) cells (i.v.; 2 × 104 B‐ALL cells/ mouse) and survival was monitored for >3 months. Once signs of leukemia manifested (the detection of GFP+ cells in the peripheral blood, lethargy, ruffled fur, labored breathing, or greater than 7% weight loss), mice were removed from the study.
Hamilton J.A., Lee M.Y., Hunter R., Ank R.S., Story J.Y., Talekar G., Sisroe T., Ballak D.B., Fedanov A., Porter C.C., Eisenmesser E.Z., Dinarello C.A., Raikar S.S., DeGregori J, & Henry C.J. (2021). Interleukin‐37 improves T‐cell‐mediated immunity and chimeric antigen receptor T‐cell therapy in aged backgrounds. Aging Cell, 20(2), e13309.
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5

CD8+ T Cell Depletion and CDK4/6i + MEKi Dosing

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To deplete CD8+ T cells, mice were treated with 300 μg of anti-CD8α (clone 53–6.72, Bio X Cell) every 3 days for the duration of the experiment, starting 2 days before tumor implantation. Mouse IgG2a antibody (clone 2A3, Bio X Cell) was used as a control. An intermittent dosing schedule was used in which mice were fed chow dosed with a combination CDK4/6i + MEKi for three weeks and then switched to chow dosed with only MEKi for one week.
Teh J.L., Erkes D.A., Cheng P.F., Tiago M., Wilski N.A., Field C.O., Chervoneva I., Levesque M.P., Xu X., Dummer R, & Aplin A.E. (2020). Activation of CD8+ T cells contributes to antitumor effects of CDK4/6 inhibitors plus MEK inhibitors. Cancer immunology research, 8(9), 1114-1121.
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