Western blot analysis of extracts from various cell lines using PGK1 Antibody (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
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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
PGK1 Antibody recognizes endogenous levels of total PGK1 protein.
Human, Mouse, Rat
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Thr298 of human PGK1 protein. Antibodies are purified by protein A and peptide affinity chromatography.
PGK1 (phosphoglycerate kinase) is an essential enzyme in the glycolysis pathway (1). It catalyzes the reversible phospho-transfer reaction from 1,3-diphosphoglycerate to ADP to form ATP and 3-phosphoglycerate. The expression of PGK1 is upregulated in many cancer types and plays an important role in cancer cell proliferation and metastasis (2-5). PGK1 can also function as a protein kinase. ERK can phosphorylate PGK1 at Ser203. This phosphorylation changes PGK1 conformation and leads to its translocation from cytoplasm to mitochondria. There it interacts and phosphorylates PDHK1 (pyruvate dehydrogenase kinase isomerase 1), which leads to PDHK1 inhibition, blocks pyruvate to coenzyme A conversion, and promotes cytosolic lactate concentration (6). Acetylated PGK1 can phosphorylate Beclin1 to induce autophagy (7). PGK1 is activated by autophosphorylation at Tyr324. PTEN can dephosphorylate PGK1 at this site to downregulate its activity. In cancer cells, loss of PTEN enhances PGK1 activity and promotes glycolysis and tumor growth (8).
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