This antibody gave a positive signal in the following lysates:
TE 671 Whole Cell, Mouse Liver Tissue, Rat Thymus Tissue, Mouse Pancreas Tissue; Jurkat Whole Cell - Staurosporine Treated (24hr, 500nM), HeLa Whole Cell - Staurosporine Treated (24hr, 500nM), HeLa Whole Cell - Bleomycin Treated (20U/ml), HeLa Whole Cell, Mouse Kidney Tissue, Rat Liver Tissue
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Use a concentration of 1 µg/ml. Detects a band of approximately 141 kDa (predicted molecular weight: 141 kDa).
Use a concentration of 5 µg/ml.
RelevanceMTR encodes the enzyme 5-methyltetrahydrofolate-homocysteine methyltransferase. This enzyme, also known as cobalamin-dependent methionine synthase, catalyzes the final step in methionine biosynthesis. Mutations in MTR have been identified as the underlying cause of methylcobalamin deficiency complementation group G.
ICC/IF image of ab66039 stained MCF7 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab66039, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 100% methanol fixed (5 min) HeLa, Hek293 and HepG2 cells at 5µg/ml, and in 4% PFA fixed (10 min) MCF7 cells at 5µg/ml.
IHC image of ab66039 staining in pancreas formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab66039, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
References for Anti-MTR antibody (ab66039)
This product has been referenced in:
Bito T et al. A dodecylamine derivative of cyanocobalamin potently inhibits the activities of cobalamin-dependent methylmalonyl-CoA mutase and methionine synthase of Caenorhabditis elegans. FEBS Open Bio4:722-9 (2014).
Read more (PubMed: 25161880) »
Pérez-Sepúlveda A et al. Levels of key enzymes of methionine-homocysteine metabolism in preeclampsia. Biomed Res Int2013:731962 (2013).
Read more (PubMed: 24024209) »