Western blot analysis of extracts from various cell lines using CMTM6 Antibody (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected (+) with constructs expressing Myc/DDK-tagged full-length human CMTM6 (CMTM6-Myc/DDK), Myc/DDK-tagged full-length mouse CMTM6 (CMTM6 mouse-Myc/DDK), Myc/DDK-tagged full-length human CMTM4 (CMTM4-Myc/DDK), Myc/DDK-tagged full-length mouse CMTM4 (CMTM4 mouse-Myc/DDK), using CMTM6 Antibody (upper), Myc-Tag (71D10) Rabbit mAb #2278 (middle), and β-actin (D6A8) Rabbit mAb #8457 (lower).
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
CMTM6 Antibody recognizes endogenous levels of total CMTM6 protein. This antibody does not cross-react with CMTM4 protein.
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ala176 of human CMTM6 protein. Antibodies are purified by protein A and peptide affinity chromatography.
CKLF-like MARVEL transmembrane domain-containing protein 6 (CMTM6) is a member of the chemokine-like factor (CKLF)-like MARVEL transmembrane domain-containing family (1). CMTM6 stabilizes plasma membrane expression of PD-L1, an immune inhibitory ligand critical for immune tolerance to self and anti-tumor immunity (2,3). CMTM6 associates with PD-L1 at recycling endosomes, where it protects PD-L1 from being targeted for lysosomal degradation by preventing STUB1-mediated PD-L1 ubiquitination (2,3). CMTM6 may stabilize PD-L1 expression on antigen presenting cells and potentiate inhibitory signaling by PD-1 on T cells, triggering T cell inhibition and exhaustion. CMTM6 has also been shown to interact with CD58, ARG1, ENO1, and TMPO (2). Due to the role of CMTM6 in regulating the immune system, it is being investigated as an immunotherapeutic target for the treatment of cancer.
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