Western blot analysis of extracts from PANC-1 cells or HeLa cells, vehicle-treated (-) or treated with carbonyl cyanide m-chlorophenylhydrazone (CCCP) (100 μM, 2 hr; +), using PGAM5 (Long Isoform) Antibody (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
|REACTIVITY||H M R Mk|
|MW (kDa)||28, 30|
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
PGAM5 (Long Isoform) Antibody recognizes endogenous levels of total PGAM5 long isoform protein. This antibody specifically recognizes the PGAM5 long isoform (UniProt #Q96HS1-1) and does not recognize the PGAM5 short isoform (UniProt #Q96HS1-2).
Human, Mouse, Rat, Monkey
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human PGAM5 long isoform (UniProt #Q96HS1-1). Antibodies are purified by protein A and peptide affinity chromatography.
Phosphoglycerate mutase family 5 (PGAM5) is a mitochondrial serine/threonine protein phosphatase (1,2). PGAM5 is required for necrosis induced by TNF-α or reactive oxygen species (2). In addition, PGAM5 dephosphorylates FUNDC1 to induce mitophagy under mitochondrial stresses (3). PGAM5 is also essential for RIPK1-mediated mitophagy activation during extracellular matrix (ECM) detachment of cancer cells (4). Furthermore, PGAM5 is cleaved by the protease presenilin-associated rhomboid-like protein (PARL) in response to mitochondrial stresses (such as induced by carbonyl cyanide m-chlorophenylhydrazone (CCCP) treatment). The cleaved PGAM5, released from the mitochondrial membrane, dephosphorylates β-catenin in the cytosol leading to its stabilization. Increased levels of β-catenin stimulate mitochondrial biogenesis (5).
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