Use 0.1µg for 106 cells. ab170191-Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.
FunctionInvolved in the regulation of homocysteine metabolism. Converts betaine and homocysteine to dimethylglycine and methionine, respectively. This reaction is also required for the irreversible oxidation of choline.
Tissue specificityFound exclusively in liver and kidney.
PathwayAmine and polyamine degradation; betaine degradation; sarcosine from betaine: step 1/2. Amino-acid biosynthesis; L-methionine biosynthesis via de novo pathway; L-methionine from L-homocysteine (BhmT route): step 1/1.
Overlay histogram showing HepG2 cells stained with ab52144 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab52144, 0.1μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H&L) (ab150113) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in HepG2 cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
Western blot - BHMT antibody [clone 3D6] (ab52144)
Anti-BHMT antibody [clone 3D6] (ab52144) at 1/1000 dilution + extracts of mouse liver at 20 µg
Secondary goat anti-mouse secondary antibody conjugated to HRP Developed using the ECL technique
Predicted band size : 45 kDa Observed band size : 45 kDa
References for Anti-BHMT antibody [clone 3D6] (ab52144)
This product has been referenced in:
Li S et al. Regulation of homocysteine homeostasis through the transcriptional coactivator PGC-1alpha. Am J Physiol Endocrinol Metab296:E543-8 (2009).
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