Western blot analysis of extracts from activated primary human CD8+ T cells and HDLM-2 cells using 4-1BB/CD137/TNFRSF9 Antibody.
Western blot analysis of extracts from COS-7 cells, mock transfected (-) or transfected with a construct expressing Myc-tagged full-length human 4-1BB/CD137/TNFRSF9 (h4-1BB/CD137/TNFRSF9-Myc; +), untreated (-) or treated (+) by PNGase F, using 4-1BB/CD137/TNFRSF9 Antibody (upper), Myc-Tag (9B11) Mouse mAb #2276 (middle), or β-Actin (D6A8) Rabbit mAb #8457 (lower).
|MW (kDa)||32, 40|
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not aliquot the antibody.
For western blots, incubate membrane with diluted primary antibody in 5% w/v nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
NOTE: Please refer to primary antibody product webpage for recommended antibody dilution.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).
NOTE: Volumes are for 10 cm x 10 cm (100 cm2) of membrane; for different sized membranes, adjust volumes accordingly.
* Avoid repeated exposure to skin.
posted June 2005
revised June 2020
Protocol Id: 263
4-1BB/CD137/TNFRSF9 Antibody recognizes endogenous levels of total 4-1BB/CD137/TNFRSF9 protein.
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gln128 of human 4-1BB/CD137/TNFRSF9 protein. Antibodies are purified by protein A and peptide affinity chromatography.
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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