Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing MycDDK-tagged full-length human HMGCS1 (Human HMGCS1 (MycDDK-tagged); +) or Myc-tagged full-length human HMGCS2 (Human HMGCS2 (Myc-tagged); +) using HMGCS2 (D8B5M) Rabbit mAb (upper), Myc-Tag (71D10) Rabbit mAb #2278 (middle), or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western blot analysis of extracts from mouse liver and rat liver using HMGCS2 (D8B5M)) Rabbit mAb.
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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
HMGCS2 (D8B5M) Rabbit mAb recognizes endogenous levels of total HMGCS2 protein. This antibody does not cross-react with HMGCS1 protein.
Human, Mouse, Rat
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ala45 of human HMGCS2 protein.
Mitochondrial 3-hydroxy-3-methylglutaryl-CoA synthase (HMGCS2) generates hydroxymethylglutaryl-CoA (HMG-CoA) from acetyl-CoA and acetoacetyl-CoA, a rate-limiting step in ketogenesis (1). Starvation or a high-fat and low-carbohydrate diet increases the levels of hepatic FGF21, which in turn up-regulates HMGCS2 expression (2). Furthermore, mTORC1 inhibition was shown to be required for the increase of HMGCS2 expression mediated by PPARα in response to fasting (3). In addition, studies on mice lacking HMGCS2 suggest that ketogenesis plays a role in the prevention of diet-induced fatty liver injury and hyperglycemia (4).
Explore pathways + proteins related to this product.