Flow cytometric analysis of human peripheral blood mononuclear cells, untreated (left) or treated with anti-CD3 plus anti-CD28 (10 μg/ml, 72 hr) (right), using 4-1BB/CD137/TNFRSF9 (D2Z4Y) Rabbit mAb (PE Conjugate) (top row) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control (PE Conjugate) #5742 (bottom row) and co-stained with a CD8 antibody.
Flow cytometric analysis of live Jurkat cells (blue, negative) and HDLM-2 cells (green, positive) using 4-1BB/CD137/TNFRSF9 (D2Z4Y) Rabbit mAb (PE Conjugate) (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control (PE Conjugate) #5742 (dashed lines).
This Cell Signaling Technology antibody is conjugated to phycoerythrin (PE) and tested in-house for direct flow cytometric analysis in human cells. This antibody is expected to exhibit the same species cross-reactivity as the unconjugated 4-1BB/CD137/TNFRSF9 (D2Z4Y) Rabbit mAb #34594.
Supplied in PBS (pH 7.2), less than 0.1% sodium azide and 2 mg/ml BSA. Store at 4°C. Do not aliquot the antibodies. Protect from light. Do not freeze.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
NOTE: When including fluorescent cellular dyes in your experiment (including viability dyes, DNA dyes, etc.), please refer to the dye product page for the recommended protocol. Visit www.cellsignal.com for a full listing of cellular dyes validated for use in flow cytometry.
NOTE: Count cells using a hemocytometer or alternative method.
NOTE: If using whole blood, lyse red blood cells and wash by centrifugation prior to Immunostaining.
NOTE: Optimal centrifugation conditions will vary depending upon cell type and reagent volume. Generally, 150-300g for 1-5 minutes will be sufficient to pellet the cells.
posted June 2017
revised June 2020
Protocol Id: 1504
4-1BB/CD137/TNFRSF9 (D2Z4Y) Rabbit mAb (PE Conjugate) recognizes endogenous levels of total 4-1BB/CD137/TNFRSF9 protein.
Monoclonal antibody is produced by immunizing animals with a recombinant protein specific to the extracellular domain of human 4-1BB/CD137/TNFRSF9 protein.
TNFRSF9 is a member of the tumor necrosis factor receptor superfamily (1, 2). It is also called 4-1BB or CD137 (1, 2). 4-1BB/CD137/TNFRSF9 is expressed in activated CD4+ and CD8+ T cells, natural killer cells and dendritic cells (2-5). The ligand 4-1BBL/CD137L/TNFSF9 on antigen presenting cells binds to 4-1BB/CD137/TNFRSF9 and costimulates the activation of T cells (5). The binding of agonistic antibodies to 4-1BB/CD137/TNFRSF9 also leads to costimulation for T cell activation (5). Studies have shown the effectiveness of targeting 4-1BB/CD137/TNFRSF9 by its agonistic antibodies in cancer immunotherapy (6).
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