Anti-PSMA Purified

Glutamate carboxypeptidase II (GCPII), also known as N-acetyl-alpha-linked acidic dipeptidase I (NAALADase I), folate hydrolase (FOLH1), and prostate-specific membrane antigen (PSMA), is an approximately 95-110 kDa type II transmembrane glycoprotein expressed in various tissues. In nervous system GCPII cleaves abundant N-acetylaspartylglutamate, which is released from neurons in a calcium-dependent manner, to N-acetylaspartate and glutamate. As immoderate glutamate concentration is neurotoxic, GCPII contributes to pathological conditions regarding e.g. Alzheimer´s disease, Huntington´s disease, epilepsy, schizophrenia, stroke or neuropathic pain and appears to be an interesting therapeutic target. In jejunum GCPII hydrolyzes pteroylpoly-gamma-glutamate to folate and glutamate, enabling folate to be absorbed by gastrointestinal tract. GCPII, which is present in a number of tissues at low levels, is overexpressed in neovasculature of most solid tumours and is a target enzyme for diagnosis and treatment of prostate cancer. Normal human prostate express more mRNA coding for a cytosolic GCPII form truncated at the N-terminus (PSM´) than mRNA for membrane-bound GCPII, and this ratio is reversed upon malignant transformation.

Product Specifications

Certification

RUO

Reactivity

Pig, Mouse, Rat, Human

Immunogen

Recombinant fragment of human GCPII (amino acids 44-750) produced in S2 cells

Target Antigen

PSMA

Clone

GCP-04

Applications

ICC, WB (QC tested), IHC-P (reported)

Concentration

1 mg/mL

Format

Purified

Buffer

Phosphate buffered saline (PBS), pH 7.4, 15 mM sodium azide

References & Citations

*Barinka C, Mlcochova P, Sacha P, Hilgert I, Majer P, Slusher BS, Horejsi V, Konvalinka J: Amino acids at the N- and C-termini of human glutamate carboxypeptidase II are required for enzymatic activity and proper folding. Eur. J. Biochem. 2004, 271: 2782-2790, URL: https://www.ncbi.nlm.nih.gov/pubmed/15206943?ordin, *Barinka C, Sacha P, Sklenar J, Man P, Bezouska K, Slusher BS, Konvalinka J: Identification of the N-glycosylation sites on glutamate carboxypeptidase II necessary for proteolytic activity. Protein Sci. 2004, 13: 1627-1635, URL: https://www.ncbi.nlm.nih.gov/pubmed/15152093?ordin, *Sacha P, Zamecnik J, Barinka C, Hlouchova K, Vicha A, Mlcochova P, Hilgert I, Eckschlager T, Konvalinka J: Expression of glutamate carboxypeptidase II in human brain. Neuroscience 2007, 144: 1361-1372, URL: https://www.ncbi.nlm.nih.gov/pubmed/17150306?ordin, *Rovenska M, Hlouchova K, Sacha P, Mlcochova P, Horak V, Zamecnik J, Barinka C, Konvalinka J: Tissue expression and enzymologic characterization of human prostate specific membrane antigen and its rat and pig orthologs. Prostate 2008, 68: 171-182 , URL: https://www.ncbi.nlm.nih.gov/pubmed/18076021?ordin, *Barinka C, Rinnova M, Sacha P, Rojas C, Majer P, Slusher BS, Konvalinka J: Substrate specificity, inhibition and enzymological analysis of recombinant human glutamate carboxypeptidase II. J. Neurochem. 2002, 80: 477-487, URL: https://www.ncbi.nlm.nih.gov/pubmed/11905994?ordin, *Tykvart J, Navrátil V, Sedlák F, Corey E, Colombatti M, Fracasso G, Koukolík F, Bařinka C, Sácha P, Konvalinka J: Comparative analysis of monoclonal antibodies against prostate-specific membrane antigen (PSMA) . Prostate. 2014 Dec;74 (16) :1674-90., URL: http://www.ncbi.nlm.nih.gov/pubmed/25262926

Other References

*Ghadge GD, Slusher BS, Bodner A, Canto MD, Wozniak K, Thomas AG, Rojas C, Tsukamoto T, Majer P, Miller RJ, Monti AL, Roos RP: Glutamate carboxypeptidase II inhibition protects motor neurons from death in familial amyotrophic lateral sclerosis models. Proc Natl Acad Sci U S A. 2003 Aug 5;100 (16) :9554-9., URL: http://www.ncbi.nlm.nih.gov/pubmed/12876198?ordina, *Mesters JR, Barinka C, Li W, Tsukamoto T, Majer P, Slusher BS, Konvalinka J, Hilgenfeld R: Structure of glutamate carboxypeptidase II, a drug target in neuronal damage and prostate cancer. EMBO J. 2006, 25: 1375-1384, URL: https://www.ncbi.nlm.nih.gov/pubmed/16467855?ordin, *Davis MI, Bennett MJ, Thomas LM, Bjorkman PJ: Crystal structure of prostate-specific membrane antigen, a tumor marker and peptidase. Proc. Natl. Acad. Sci. USA 2005, 102: 5981-5986, URL: https://www.ncbi.nlm.nih.gov/pubmed/15837926?ordin

Storage Conditions

Store at 2-8°C. Do not freeze.

Specificity

The mouse monoclonal antibody GCP-04 recognizes amino acids 100-104 of extracellular domain of denaturated glutamate carboxypeptidase II (PSMA, NAALADase, FOLH1), an approximately 95-110 kDa transmembrane glycoprotein.

Applications Notes

Western blotting: Recommended dilution: 1 μg/ml; positive control: LNCaP cell line. Sample preparation: Resuspend approx. 50 mil. cells in 1 mL cold lysis buffer (1% NP-40) . Incubate 30 min on ice. Mix lysate with non-reducing/reducing Laemmli SDS-PAGE sample buffer. Both reducing and non-reducing conditions.