1/50 - 1/100. Heat up to 98 °C, below boiling, and then let cool for 10-20 min.
Application notesIs unsuitable for Flow Cyt or ICC.
FunctionConverts guanine to guanosine monophosphate, and hypoxanthine to inosine monophosphate. Transfers the 5-phosphoribosyl group from 5-phosphoribosylpyrophosphate onto the purine. Plays a central role in the generation of purine nucleotides through the purine salvage pathway.
PathwayPurine metabolism; IMP biosynthesis via salvage pathway; IMP from hypoxanthine: step 1/1.
Involvement in diseaseDefects in HPRT1 are the cause of Lesch-Nyhan syndrome (LNS) [MIM:300322]. LNS is characterized by complete lack of enzymatic activity that results in hyperuricemia, choreoathetosis, mental retardation, and compulsive self-mutilation. Defects in HPRT1 are the cause of gout HPRT-related (GOUT-HPRT) [MIM:300323]; also known as HPRT-related gout or Kelley-Seegmiller syndrome. Gout is characterized by partial enzyme activity and hyperuricemia.
Sequence similaritiesBelongs to the purine/pyrimidine phosphoribosyltransferase family.
Western blot - Anti-HPRT antibody [EPR5299] (ab109021)
Predicted band size : 25 kDa
Lane 1: Wild type HAP1 whole cell lysate (20 µg) Lane 2: HPRT1 knockout HAP1 whole cell lysate (20 µg) Lane 3: HeLa whole cell lysate (20 µg) Lane 4: A431 whole cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab109021 observed at 25 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab109021 was shown to specifically react with HPRT1 when HPRT1 knockout samples were used. Wild-type and HPRT1 knockout samples were subjected to SDS-PAGE. ab109021 and ab8245 (Mouse anti GAPDH loading control) were both diluted at 1/10000 and incubated overnight at 4°C. Blots were developed with 800CW Goat anti Rabbit and 680CW Goat anti Mouse secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
Western blot - HPRT antibody [EPR5299] (ab109021)
All lanes : Anti-HPRT antibody [EPR5299] (ab109021) at 1/10000 dilution
Lane 1 : HeLa cell lysate Lane 2 : HepG2 cell lysate Lane 3 : A549 cell lysate Lane 4 : MCF7 cell lysate