Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length human Tapasin protein (hTapasin-Myc/DDK; +), using Tapasin Antibody (upper), DYKDDDDK Tag Antibody #2368 (middle), and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western blot analysis of extracts from HeLa and B16-F10 cells, untreated (-) or treated with IFN-γ (100 ng/ml, 16 hr; +), using Tapasin Antibody (upper) or 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 BSA, 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.
From sample preparation to detection, the reagents you need for your Western Blot are now in one convenient kit: #12957 Western Blotting Application Solutions Kit
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: 10
Tapasin Antibody recognizes endogenous levels of total Tapasin protein.
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of human Tapasin protein. Antibodies are purified by peptide affinity chromatography.
TAP-associated glycoprotein (Tapasin/TAPBP) is a type-I transmembrane ER glycoprotein encoded by an MHC-linked gene. Tapasin plays a critical role in the MHC class I-restricted antigen processing and presentation pathway. Research studies have demonstrated that Tapasin is an IFNγ-inducible component of the TAP-1/2 peptide transporter complex and, within the ER lumen, facilitates the optimization of the peptide repertoire that is loaded onto MHC class I molecules (1-5). Downregulation of Tapasin and MHC class I expression has been observed in human tumors and serves as a prominent mechanism of tumor immune escape (6-9).
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