Western blot analysis of extracts from various cell lines using PCK1 (D12F5) Rabbit mAb.
Western blot analysis of extracts from 293 cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length human PCK1 (hPCK1-Myc/DDK; +), using PCK1 (D12F5) Rabbit mAb (upper), DYKDDDDK Tag (9A3) Mouse mAb #8146 (middle), or 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, 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 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: 263
PCK1 (D12F5) Rabbit mAb recognizes endogenous levels of total PCK1 protein.
Human, Mouse, Rat, Monkey
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human PCK1 protein.
Phosphoenolpyruvate carboxykinase 1 (PCK1, PEPCK1 or PEPCK-C) is a cytosolic enzyme responsible for the conversion of oxaloacetate to phosphoenolpyruvate (1). PCK1 is involved in controlling the rate-limiting step of gluconeogenesis in the liver, which generates glucose from non-carbohydrate substrates such as lactate and glycerol (2). The deacetylase SirT1 stimulates transcription of PCK1 and glucose-6-phosphatase to activate gluconeogenesis (3). Depending on nutritional state, Stat3 can inhibit PCK1 and glucose-6-phosphatase expression and suppress gluconeogenesis (4). Relatively high glucose concentration can result in acetylation of PCK1 by P300 acetyltransferase, promoting an interaction between PCK1 and the E3 ligase UBR5 that leads to the PCK1 destabilization (5).
Explore pathways + proteins related to this product.
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