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Recombinant Human PSMA/FOLH1 Protein, CF

R&D Systems | Catalog # 4234-ZN

Analyzed by SEC-MALS
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Key Product Details

  • R&D Systems CHO-derived Recombinant Human PSMA/FOLH1 Protein (4234-ZN)
  • Quality control testing to verify active proteins with lot specific assays by in-house scientists
  • All R&D Systems proteins are covered with a 100% guarantee

Source

CHO

Accession Number

Q04609

Applications

Enzyme Activity
View all PSMA/FOLH1/NAALADase I Proteins and Enzymes »

Product Specifications

Source

Chinese Hamster Ovary cell line, CHO-derived human PSMA/FOLH1/NAALADase I protein
Lys44-Ala750, with an N-terminal 6-His tag

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<1.0 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

His

Predicted Molecular Mass

80 kDa

SDS-PAGE

110 kDa, reducing conditions

Activity

Measured by its ability to hydrolyze the substrate N-acetyl-L-Asp-L-Glu into N-acetyl-L-Asp and L-Glu. The L-Glu product is measured by fluorescence after its derivatization by ortho-phthaldialdehyde.
The specific activity is >400 pmol/min/µg, as measured under the described conditions.

Reviewed Applications

Read 2 reviews rated 5 using 4234-ZN in the following applications:

Scientific Data Images for Recombinant Human PSMA/FOLH1 Protein, CF

Recombinant Human PSMA/FOLH1/NAALADase I SEC-MALS.

Recombinant Human PSMA/FOLH1/NAALADase I (Catalog # 4234-ZN) has a molecular weight (MW) of 185-204 kDa as analyzed by SEC-MALS, suggesting that this protein is a homodimer. MW may differ from predicted MW due to post-translational modifications (PTMs) present (i.e. Glycosylation).

Formulation, Preparation, and Storage

4234-ZN
Formulation Supplied as a 0.2 μm filtered solution in MES and NaCl.
Shipping The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.

Background: PSMA/FOLH1/NAALADase I

Human prostate-specific membrane antigen (PSMA), a tumor marker in prostate cancer encoded by the FOLH1 gene, is a type II transmembrane zinc metallopeptidase that is most highly expressed in the nervous system, prostate, kidney, and small intestine (1, 2). The enzyme is also known as glutamate carboxypeptidase II (GCPII), folate hydrolase 1, folypoly-gamma-glutamate carboxypeptidase (FGCP), and N-acetylated-alpha-linked acidic dipeptidase I (NAALADase I). In the brain, PSMA hydrolyzes the neurotransmitter N-acetyl-Asp-Glu to produce glutamate, another neurotransmitter. Inhibition of brain PSMA activity is considered to be a promising approach for the treatment of neurological disorders associated with glutamate excitotoxicity, such as stroke, chronic pain, and amyotrophic lateral sclerosis (3). Intestinal PSMA hydrolyzes folylpoly-gamma -glutamates, facilitating the uptake of folate (4).

References

  1. Silver, D.A. et al. (1997) Clin. Cancer Res. 3:81.
  2. Carter, R.E. et al. (1996) Pro. Natl. Acad. Sci. USA. 93:749.
  3. Jackson, P.F. and Slusher, B.S. (2001) Curr. Med. Chem. 8:949.
  4. Heston, W.D. (1997) Urology 49:104.

Long Name

Prostate-specific Membrane Antigen

Alternate Names

FGCP, FOLH1, GCP2, GCPII, mopsm, NAALAD1, NAALADase I

Entrez Gene IDs

2346 (Human); 53320 (Mouse); 85309 (Rat); 101866140 (Cynomolgus Monkey)

Gene Symbol

FOLH1

UniProt

Q04609

Additional PSMA/FOLH1/NAALADase I Products

Product Documents for Recombinant Human PSMA/FOLH1 Protein, CF

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Recombinant Human PSMA/FOLH1 Protein, CF

For research use only

Citations for Recombinant Human PSMA/FOLH1 Protein, CF

Customer Reviews for Recombinant Human PSMA/FOLH1 Protein, CF (2)

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  • Recombinant Human PSMA/FOLH1 Protein, CF
    Name: Jenna Nguyen
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Protocols

View specific protocols for Recombinant Human PSMA/FOLH1 Protein, CF (4234-ZN):

Materials
  • Assay Buffer: 50 mM HEPES, 0.1 M NaCl, pH 7.5
  • OPA Buffer: 0.2 M NaOH containing 0.1% beta -Mercaptoethanol (v/v)
  • Recombinant Human PSMA/FOLH1/NAALADase1 (rhPSMA) (Catalog # 4234-ZN)
  • Substrate: Ac-Asp-Glu  (Sigma, Catalog # A5930), 10 mM stock in 40 mM NaOH
  • ortho-phthaldialdehyde (OPA) (Sigma, Catalog # P0657), 50 mg/mL stock in DMSO
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rhPSMA to 0.4 µg/mL in Assay Buffer.
  2. Dilute Substrate to 40 µM with Assay Buffer.
  3. Combine 125 µL of 0.4 µg/mL rhPSMA and 125 µL of 40 µM Substrate. For a control, inactivate 125 µL of 0.4 µg/mL rhPSMA by heating it at 95 °C for 5 minutes, then add 125 µL of 40 µM Substrate Substrate.
  4. Incubate at 37 °C for 1 hour.
  5. Stop the reaction by heating at 95 °C for 5 minutes, then cool to room temperature.
  6. Prepare a 15 mM OPA solution in OPA Buffer.
  7. Add 250 µL of OPA solution to all vials and vortex.
  8. Incubate at room temperature for 10 minutes.
  9. Load 200 µL of reaction and control to plate.
  10. Read at excitation and emission wavelengths of 330 nm and 450 nm (top read), respectively in endpoint mode.
  11. Calculate specific activity:

     Specific Activity (pmol/min/µg) =

 Adjusted Fluorescence* (RFU) x Conversion Factor** (pmol/RFU)
Incubation time (min) x amount of enzyme (µg)

     *Adjusted for Substrate Blank

     **Derived using calibration standard L-Glutamic Acid (Sigma, Catalog # G8415).

Per Well:
  • rhPSMA: 0.020 µg
  • Substrate: 10 µM
  • OPA: 7.5 mM

FAQs

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