Western blot analysis of extracts from NIH/3T3 and HeLa cells using MEK1/2 (D1A5) Rabbit mAb (HRP Conjugate).
|REACTIVITY||H M R Mk Dm|
This Cell Signaling Technology antibody is conjugated to the carbohydrate groups of horseradish peroxidase (HRP) via its amine groups. The HRP conjugated antibody is expected to exhibit the same species cross-reactivity as the unconjugated antibody MEK1/2 (D1A5) Rabbit mAb #8727.
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 antibodies.
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: 11
MEK1/2 (D1A5) Rabbit mAb (HRP Conjugate) recognizes endogenous levels of total MEK1 and MEK2 proteins. This antibody is predicted to cross-react with MEK1/MEK2 orthologs in a variety of species.
Human, Mouse, Rat, Monkey, D. melanogaster
Hamster, Xenopus, Zebrafish, Bovine, Dog, Pig, C. elegans
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ala220 of human MEK1 protein.
MEK1 and MEK2, also called MAPK or Erk kinases, are dual-specificity protein kinases that function in a mitogen activated protein kinase cascade controlling cell growth and differentiation (1-3). Activation of MEK1 and MEK2 occurs through phosphorylation of two serine residues at positions 217 and 221, located in the activation loop of subdomain VIII, by Raf-like molecules. MEK1/2 is activated by a wide variety of growth factors and cytokines and also by membrane depolarization and calcium influx (1-4). Constitutively active forms of MEK1/2 are sufficient for the transformation of NIH/3T3 cells or the differentiation of PC-12 cells (4). MEK activates p44 and p42 MAP kinase by phosphorylating both threonine and tyrosine residues at sites located within the activation loop of kinase subdomain VIII.
Explore pathways + proteins related to this product.
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