categories: Neurobiology (Human), Neurobiology (Rodent), Neurobiology (Veterinary)

Mouse Monoclonal to GCPII / PSMA

GCP-04 (IgG1)

	Product No.	Form	Quantity	Stock	Price	Datasheet	Shop
	11-532-C025	purified	0.025 mg		choose region
	11-532-C100	purified	0.1 mg		choose region

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Background

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.

Entrez Gene (human): 11p11.2

Specificity

The 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.

Immunogen

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

Species Reactivity: Human Porcine Rat Other not determined Negative Species:	Applications: 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. Application note: Both reducing and non-reducing conditions. Immunohistochemistry (paraffin sections) Immunocytochemistry Usage note: Indicated dilutions are recommended starting points for use of this product. Working concentrations should be determined by the investigator.	General 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. [Abstract] [Full Text] 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 [Abstract] [Full Text] 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 [Abstract] [Full Text] Product Specific References 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 [Abstract] 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 [Abstract] 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 [Abstract] [Full Text] 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 [Abstract] 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 [Abstract]

Species Reactivity:

Human
Porcine
Rat
Other not determined

Negative Species:

Applications:

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.
Application note: Both reducing and non-reducing conditions.
Immunohistochemistry (paraffin sections)
Immunocytochemistry

Usage note:
Indicated dilutions are recommended starting points for use of this product. Working concentrations should be determined by the investigator.

General 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. [Abstract] [Full Text]

*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 [Abstract] [Full Text]

*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 [Abstract] [Full Text]

Product Specific References

*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 [Abstract]

*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 [Abstract]

*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 [Abstract] [Full Text]

*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 [Abstract]

*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 [Abstract]

For research use only. Not for drug, diagnostic or other use.

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Example Data

Fig. 1.

Fig. 1. Immunostaining of a fragment of human GCPII (aminoacids 44-750) produced in S2 cells on Western blot by GCP-04 monoclonal antibody. Lanes 1, 2, 3 represent 800, 400 and 200 pg of the protein.

Fig. 2.

Fig. 2. Immunohistochemistry of GCPII in human Medulla oblongata by GCP-04 monoclonal antibody. Mag. 40x; positive astrocytes in white matter.

Fig. 3.

Fig. 3. Immunohistochemistry of GCPII in human prostate by GCP-04 monoclonal antibody. Mag. 400x; positive epithelium of the prostate glands.

Fig. 4.

Fig. 4. Immunohistochemistry of GCPII in porcine kidney by GCP-04 monoclonal antibody. Highly positive proximal glomeruli.

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