Western blot analysis of total cell lysates from P3HR-1 and Ramos cells using AID (EK2 5G9) Rat mAb.
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 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.
* Avoid repeated exposure to skin.
posted June 2005
revised June 2020
Protocol Id: 340
AID (EK2 5G9) Rat mAb detects endogenous levels of total AID protein.
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to a region surrounding Asp191 of human AID.
Activation-induced cytidine deaminase (AID) is thought to modify RNA due to its high homology to the RNA editing enzyme APOBEC-1. This function, however, has not been confirmed in in vitro studies, which show that AID has significant cytidine deaminase activity, and that this activity is blocked by zinc chelation (1).
The B cell immune system must specifically recognize several infectious agents, which vastly outnumber immunoglobulin gene segments present in a given organism. Mechanisms such as somatic hypermutation, isotype switch recombination and gene conversion introduce diversity and specificity to the immune system. Analysis of mouse models and patients with AID deficiency has established a link between all three of these mechanisms and AID function (2). AID protein is detected in germinal center centroblast and germinal center derived lymphomas (Burkitt lymphoma), but not in pre-germinal center B cells or post-germinal center neoplasms (B cell chronic lymphocytic leukemia and multiple myeloma) (3).
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