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Cd28 cd49d mab

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The CD28/CD49d mAb is a monoclonal antibody that recognizes the CD28 and CD49d cell surface molecules. It is commonly used in research applications to stimulate T-cell activation and proliferation.

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Lab products found in correlation

Perm wash solution, by BD (3 mentions) Brefeldin a, by BD (3 mentions) Monensin, by BD (3 mentions) Anti ifn γ mab, by BD (2 mentions)

Close spelling variants of this product

Anti-CD28 and anti-CD49d mAbs Anti-CD28 and anti-CD49d monoclonal antibodies (mAb) CD28/CD49d

3 protocols using cd28 cd49d mab

1

Characterization of Anti-Tumor CD8+ T Cells

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CD107a degranulation and IFN-γ producing CD3+CD8+ T cells were identified within SMM MP-CTL by flow cytometry. Briefly, SMM MP-CTL were stimulated with HLA-A2+ or HLA-A2- MM cell lines, K562 cells, K562-A*0201 cells pulsed with respective peptide or K562-A*0201 cells alone in the presence of CD107a anti-human mAb. SMM MP-CTL alone served as a negative control. After 1 hour incubation, CD28/CD49d mAb (BD), as well as protein transport inhibitors Brefeldin A and Monensin (BD), were added for an additional 5 hours. Cells were harvested, washed in FACS buffer, and incubated with mAbs specific to CD3, CD8, CCR7, CD45RO, CD69 and/or CD137 antigens. After surface staining, cells were washed, fixed/permeabilized, stained with anti-IFN-γ mAb (BD), washed with Perm/Wash solution (BD), fixed in 2% paraformaldehyde, and analyzed by flow cytometry.
Bae J., Prabhala R., Voskertchian A., Brown A., Maguire C., Richardson P., Dranoff G., Anderson K.C, & Munshi N.C. (2014). A Multiepitope of XBP1, CD138 and CS1 Peptides Induces Myeloma-Specific Cytotoxic T lymphocytes in T cells of Smoldering Myeloma Patients. Leukemia, 29(1), 218-229.
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2

Characterization of Anti-Tumor CD8+ T Cells

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CD107a degranulation and IFN-γ producing CD3+CD8+ T cells were identified within SMM MP-CTL by flow cytometry. Briefly, SMM MP-CTL were stimulated with HLA-A2+ or HLA-A2- MM cell lines, K562 cells, K562-A*0201 cells pulsed with respective peptide or K562-A*0201 cells alone in the presence of CD107a anti-human mAb. SMM MP-CTL alone served as a negative control. After 1 hour incubation, CD28/CD49d mAb (BD), as well as protein transport inhibitors Brefeldin A and Monensin (BD), were added for an additional 5 hours. Cells were harvested, washed in FACS buffer, and incubated with mAbs specific to CD3, CD8, CCR7, CD45RO, CD69 and/or CD137 antigens. After surface staining, cells were washed, fixed/permeabilized, stained with anti-IFN-γ mAb (BD), washed with Perm/Wash solution (BD), fixed in 2% paraformaldehyde, and analyzed by flow cytometry.
Bae J., Prabhala R., Voskertchian A., Brown A., Maguire C., Richardson P., Dranoff G., Anderson K.C, & Munshi N.C. (2014). A Multiepitope of XBP1, CD138 and CS1 Peptides Induces Myeloma-Specific Cytotoxic T lymphocytes in T cells of Smoldering Myeloma Patients. Leukemia, 29(1), 218-229.
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3

Functional Evaluation of BCMA-CTL

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The functional immune activities of BCMA-CTL were measured by CD107a degranulation and Th1-specific cytokines production by flow cytometry. In brief, BCMA-CTL were co-incubated with tumor cells or peptide loaded T2 cells in the presence of CD107a mAb. After 1-hour incubation, CD28/CD49d mAb, Brefeldin A, and Monensin (BD) were added for an additional 5 hours. Cells were harvested, washed in PBS, and incubated with mAbs specific to T cell antigens. After surface staining, cells were washed, fixed/permeabilized, stained with anti-IFN-γ/IL-2/TNF-α mAbs, washed with Perm/Wash solution (BD), fixed in 2% paraformaldehyde, and analyzed by flow cytometry.
Bae J., Samur M., Richardson P., Munshi N.C, & Anderson K.C. (2019). Selective Targeting of Multiple Myeloma by B cell Maturation Antigen (BCMA)-specific Central Memory CD8+ Cytotoxic T Lymphocytes: Immunotherapeutic Application in Vaccination and Adoptive Immunotherapy. Leukemia, 33(9), 2208-2226.
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