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Anti cd49d clone l25

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Anti-CD49d clone L25 is a monoclonal antibody that binds to the CD49d antigen. CD49d is a subunit of the VLA-4 integrin complex. The antibody can be used for the identification and analysis of CD49d-expressing cells in various applications.

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Lab products found in correlation

Fetal bovine serum (fbs), by Avantor (2 mentions) Brefeldin a solution, by Thermo Fisher Scientific (2 mentions) Anti cd28 clone l293, by BD (2 mentions) Pepmix sars cov 2 peptide spike glycoprotein, by JPT Peptide Technologies (2 mentions) Pepmix sars cov 2 peptide ncap, by JPT Peptide Technologies (2 mentions) Penicillin, by Thermo Fisher Scientific (2 mentions) Streptomycin, by Thermo Fisher Scientific (2 mentions) Rpmi 1640 medium, by Corning (1 mentions) Rpmi 1640, by Corning (1 mentions) V bottom plate, by Corning (1 mentions) Anti cd107a pe cf594 clone h4a3, by BD (1 mentions) Brefeldin a, by Merck Group (1 mentions) Monensin, by BD (1 mentions)

Close spelling variants of this product

CD49d (clone L25) Anti-CD49d (L25 CD49d (L25) Anti-CD49d

3 protocols using anti cd49d clone l25

1

SARS-CoV-2 Antigen-Specific T Cell Profiling

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Cryopreserved PBMCs were thawed and cultured overnight. Baseline specimens were stained directly in the absence of ex vivo stimulation. For identification of antigen-specific T cells through intracellular cytokine staining, 6×106 cells were stimulated with 0.5 mg/ml anti-CD49d clone L25 (BD Biosciences) and 0.5 mg/ml anti-CD28 clone L293 (BD Biosciences) in the absence or presence of peptides for 4h in RP10 media (RPMI 1640 medium (Corning) supplemented with 10% fetal bovine serum (FBS, VWR), 1% penicillin (GIBCO), and 1% streptomycin (GIBCO)) in the presence of 3 mg/ml Brefeldin A Solution (eBioscience). Peptides used were 1 mM PepMix SARS-CoV-2 Peptide (Spike Glycoprotein) (JPT Peptide Technologies), 1 mM PepMix SARS-CoV-2 Peptide (NCAP) (JPT Peptide Technologies), or 1 mM PepMix SARS-CoV-2 Peptide (MEVTPSGTWL) (custom synthesized by JPT Peptide Technologies).
Ma T., Ryu H., McGregor M., Babcock B., Neidleman J., Xie G., George A.F., Frouard J., Murray V., Gill G., Ghosn E., Newell E., Lee S, & Roan N.R. (2021). Protracted yet coordinated differentiation of long-lived SARS-CoV-2-specific CD8+ T cells during COVID-19 convalescence. bioRxiv.
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2

SARS-CoV-2 Antigen-Specific T Cell Assay

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Cryopreserved PBMCs were thawed and cultured overnight. Baseline specimens were stained directly in the absence of ex vivo stimulation. For identification of Ag-specific T cells through intracellular cytokine staining, 6 × 106 cells were stimulated with 0.5 mg/ml anti-CD49d clone L25 (BD Biosciences) and 0.5 mg/ml anti-CD28 clone L293 (BD Biosciences) in the absence or presence of peptides for 4 h in RP10 media (RPMI 1640 [Corning] supplemented with 10% FBS [VWR International], 1% penicillin [Life Technologies], and 1% streptomycin [Life Technologies]) in the presence of 3 μg/ml brefeldin A solution (eBioscience). The peptides used were 1 mM PepMix SARS-CoV-2 peptide (spike glycoprotein) (JPT Peptide Technologies), 1 mM PepMix SARS-CoV-2 Peptide (NCAP) (JPT Peptide Technologies), or 1 mM PepMix SARS-CoV-2 Peptide (MEVTPSGTWL) (custom synthesized by JPT Peptide Technologies).
Ma T., Ryu H., McGregor M., Babcock B., Neidleman J., Xie G., George A.F., Frouard J., Murray V., Gill G., Ghosn E., Newell E.W., Lee S.A, & Roan N.R. (2021). Protracted yet Coordinated Differentiation of Long-Lived SARS-CoV-2-Specific CD8+ T Cells during Convalescence. Journal of immunology (Baltimore, Md. : 1950), 207(5), 1344-1356.
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3

Stimulating and Analyzing PBMCs

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PBMCs were seeded in complete medium at 2 x 10 6 cells/mL in 96-well V-bottom plates (Corning) with unconjugated anti-CD28 (clone L293) and anti-CD49d (clone L25) (each at 3 µL/mL; BD Biosciences), brefeldin A (1 µL/mL; Sigma-Aldrich), monensin (0.7 µL/mL; BD Biosciences), anti-CD107a-PE-CF594 (clone H4A3; BD Biosciences), and the relevant viral peptides (each at a final concentration of 0.5 µg/mL). Unstimulated negative controls were included in each assay. Cells were analyzed by flow cytometry after incubation for 6 hours at 37°C.
Sekine T., Perez-Potti A., Nguyen S., Gorin J.B., Wu V.H., Gostick E., Llewellyn-Lacey S., Hammer Q., Falck-Jones S., Vangeti S., Yu M., Smed-Sörensen A., Gaballa A., Uhlin M., Sandberg J.K., Brander C., Nowak P., Goepfert P.A., Price D.A., Betts M.R, & Buggert M. (2020). TOX is expressed by exhausted and polyfunctional human effector memory CD8+ T cells. Science immunology, 5(49).
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