Western blot analysis of extracts from NK-92 cells, untreated (-) or treated as indicated with TPA (12-O-Tetradecanoylphorbol-13-Acetate) #4174 (80 nM, 5 hr; +) and Ionomycin, Calcium Salt #9995 (3 μM, 5 hr; +), or Brefeldin A #9972 (300 ng/mL, last 4 hr of stimulation; +), using XCL1/XCL2 Antibody (upper) or 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 XCL1 protein (hXCL1-Myc/DDK; +) and Myc/DDK-tagged full-length human XCL2 protein (hXCL2-Myc/DDK; +), using XCL1/XCL2 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
XCL1/XCL2 Antibody recognizes endogenous levels of total XCL1 and XCL2 proteins.
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro99 of human XCL1 protein. Antibodies are purified by peptide affinity chromatography.
XCL1 (Lymphotactin, SCM-1 alpha, Chemokine (C motif) ligand 1) and XCL2 (SCM-1 beta, Chemokine (C motif) ligand 2) comprise the C chemokine family, lacking two of the four invariant cysteines normally found on chemokines (1,4). Their sequences differ by 2 amino acids and they are primarily expressed by CD8+ T cells and NK cells (1-5). XCL1 and XCL2 act as ligands for the chemokine receptor XCR1 (1-5). XCR1-positive dendritic cells cross-present antigens to naïve CD8+ T cells, priming them to become activated cytotoxic CD8+ T cells (6-7). In addition, in mouse models, the XCL1-XCR1 signaling axis was shown to be involved in the formation of self-tolerance through the development of Treg cells within the thymus (8).
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