Western blot analysis of extracts from Caki-1 and Hep G2 cells using CXCL1/CXCL2 (E5M6D) Rabbit mAb (Biotinylated) (upper) or β-Actin (8H10D10) Mouse mAb #3700 (lower).
This Cell Signaling Technology antibody is conjugated to biotin under optimal conditions. The biotinylated antibody is expected to exhibit the same species cross-reactivity as the unconjugated CXCL1/CXCL2 (E5M6D) Rabbit mAb #24376.
Supplied in 136 mM NaCl, 2.6 mM KCI, 12 mM sodium phosphate (pH 7.4) dibasic, 2 mg/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.
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.
Do not add Anti-biotin, HRP-linked Antibody for detection of biotinylated protein markers. There is no need. The Streptavidin-HRP will also visualize the biotinylated markers.
* Avoid repeated exposure to skin.
posted June 2005
revised June 2020
Protocol Id: 266
CXCL1/CXCL2 (E5M6D) Rabbit mAb (Biotinylated) recognizes endogenous levels of total CXCL1 and CXCL2 proteins. This antibody is more sensitive for detection of CXCL1 protein than CXCL2 protein.
Monoclonal antibody is produced by immunizing animals with human CXCL1 recombinant protein.
CXCL1 and CXCL2 (GRO-α and GRO-β) are two of the chemokines that act as ligands for the chemokine receptor CXCR2 (1). CXCR2 is expressed by several types of leukocytes and plays an important role in the recruitment of neutrophils to sites of inflammation (2). Although neutrophil recruitment is an important component of the innate immune system’s response to infection or injury, prolonged or excessive neutrophil recruitment can lead to inflammatory disease. Therefore, blocking CXCR2 has been pursued as a therapeutic approach for a variety of inflammatory diseases (3). In addition, CXCL1 and CXCL2 expression can be elevated in the tumor microenvironment, which contributes to tumor cell survival through enhanced angiogenesis and recruitment of immunosuppressive myeloid-derived suppressor cells (MDSCs) (4,5).
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