The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 - 5 µg/ml. Predicted molecular weight: 57 kDa.
Use a concentration of 10 µg/ml.
Use a concentration of 5 µg/ml.
Use 1µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
Use at an assay dependent concentration.
PathwayAmino-acid biosynthesis; L-serine biosynthesis; L-serine from 3-phospho-D-glycerate: step 1/3.
Involvement in diseaseDefects in PHGDH are the cause of phosphoglycerate dehydrogenase deficiency (PHGDH deficiency) [MIM:601815]. It is characterized by congenital microcephaly, psychomotor retardation, and seizures.
Sequence similaritiesBelongs to the D-isomer specific 2-hydroxyacid dehydrogenase family.
PHGDH was immunoprecipitated using 0.5mg Jurkat whole cell extract, 10µg of Mouse monoclonal to PHGDH and 50µl of protein G magnetic beads (+). No antibody was added to the control (-). The antibody was incubated under agitation with Protein G beads for 10min, Jurkat whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation. Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab57030. Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/5000 dilution. Band: 57kDa; PHGDH.
Western blot - PHGDH antibody (ab57030)
Predicted band size : 57 kDa
Flow Cytometry - Anti-PHGDH antibody (ab57030)
Overlay histogram showing HeLa cells stained with ab57030 (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 (ab57030, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
References for Anti-PHGDH antibody (ab57030)
This product has been referenced in:
Kim SK et al. Differential expression of enzymes associated with serine/glycine metabolism in different breast cancer subtypes. PLoS One9:e101004 (2014).
Read more (PubMed: 24979213) »
Bollig-Fischer A et al. Oncogene activation induces metabolic transformation resulting in insulin-independence in human breast cancer cells. PLoS One6:e17959 (2011).
Read more (PubMed: 21437235) »