Western blot analysis of extracts from various cell lines using hnRNP A2/B1 (2A2) Mouse mAb.
|MW (kDa)||34 to 36|
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. 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 datasheet or 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 November 2013
Reprobing of an existing membrane is a convenient means to immunoblot for multiple proteins independently when only a limited amount of sample is available. It should be noted that for the best possible results a fresh blot is always recommended. Reprobing can be a valuable method but with each reprobing of a blot there is potential for increased background signal. Additionally, it is recommended that you verify the removal of the first antibody complex prior to reprobing so that signal attributed to binding of the new antibody is not leftover signal from the first immunoblotting experiment. This can be done by re-exposing the blot to ECL reagents and making sure there is no signal prior to adding the next primary antibody.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalently purified water.
posted June 2005
revised June 2016
Protocol Id: 19
hnRNP A2/B1 (2A2) Mouse mAb detects endogenous levels of total hnRNP A2/B1 proteins. This antibody does not cross-react with other proteins.Species Reactivity:
Monoclonal antibody is produced by immunizing animals with a human hnRNP B1 protein.
Heterogeneous nuclear ribonucleoprotein A2/B1 (hnRNP A2/B1) is a member of the hnRNP A/B family of related RNA binding proteins that bind pre-mRNA and are involved in the processing, metabolism and transport of nuclear pre-mRNA transcripts (1). Alternative splicing produces transcripts that encode two homologous hnRNP proteins, hnRNPA2 and hnRNPB1, from a single gene sequence (2). Studies demonstrate hnRNP A2/B1 splicing repression across multiple targets (3,4) and that both proteins can bind and protect telomere repeat sequences from DNase digestion (5,6). Altered expression of hnRNP B1 is seen in several forms of cancer, including squamous cell carcinoma, adenocarcinoma, and various forms of lung cancer (7). Over expression of hnRNP B1 may be associated with inhibition of DNA-PK activity and impaired DNA repair during early stages of cancer development (8). Autoantigens to hnRNP A2/B1 (termed RA33) are associated with rheumatoid arthritis, systemic lupus erythromatosus and mixed connective tissue disease (9-11).
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