Western blot analysis of extracts from various cell lines using IL3RA/CD123 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
IL3RA/CD123 Antibody recognizes endogenous levels of total IL3RA/CD123 protein.
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of human IL3RA/CD123 protein. Antibodies are purified by peptide affinity chromatography.
IL3RA/CD123 is the alpha subunit of an IL-3 receptor that also contains the common beta chain (βc) shared with receptors for other cytokines. IL3RA/CD123 alone binds to IL-3 with high specificity but low affinity and it does not transduce intracellular signaling, while βc alone does not bind to IL-3 appreciably. Together, IL3RA/CD123 and βc form the functional high-affinity receptor for IL-3. IL3RA/CD123 is mainly expressed on the hematopoietic progenitor cells, primarily the granulocytic and monocytic linages (1,2). IL3RA/CD123 is highly expressed on leukemic stem cells in the majority of AML (acute myeloid leukemia) patients and is correlated with poor prognosis. In addition, overexpression of IL3RA/CD123 has also been reported for other hematopoietic malignancies (3-5). It is considered as a valuable therapeutic target for these malignancies (6-8).
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