Recombinant Human PSMA/FOLH1 Protein, CF

Catalog # Availability Size / Price Qty
4234-ZN-010
R&D Systems Recombinant Proteins and Enzymes
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Product Details
Citations (7)
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Reviews (2)

Recombinant Human PSMA/FOLH1 Protein, CF Summary

Product Specifications

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.
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.
Source
Chinese Hamster Ovary cell line, CHO-derived human PSMA/FOLH1/NAALADase I protein
Lys44-Ala750, with an N-terminal 6-His tag
Accession #
N-terminal Sequence
Analysis
His
Predicted Molecular Mass
80 kDa
SDS-PAGE
110 kDa, reducing conditions

Product Datasheets

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4234-ZN

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

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.

Assay Procedure

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
Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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
Entrez Gene IDs
2346 (Human); 53320 (Mouse); 85309 (Rat); 101866140 (Cynomolgus Monkey)
Alternate Names
Cell growth-inhibiting gene 27 protein; cell growth-inhibiting protein 27; EC 3.4.17.21; FGCP; folate hydrolase (prostate-specific membrane antigen) 1; Folate hydrolase 1; FOLH1; FOLHNAALADase I; Folylpoly-gamma-glutamate carboxypeptidase; GCP2; GCPII; GCPIImGCP; glutamate carboxylase II; glutamate carboxypeptidase 2; Glutamate carboxypeptidase II; Membrane glutamate carboxypeptidase; mopsm; NAALAD1; NAALAD1N-acetylated-alpha-linked acidic dipeptidase I; NAALADase I; NAALAdase; N-acetylated alpha-linked acidic dipeptidase 1; prostate specific membrane antigen variant F; Prostate-specific membrane antigen; PSMA; PSMAFGCP; PSMPteroylpoly-gamma-glutamate carboxypeptidase

Citations for Recombinant Human PSMA/FOLH1 Protein, CF

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

7 Citations: Showing 1 - 7
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  1. Prostate-Specific Membrane Antigen and Esterase Dual Responsive Camptothecin-Oligopeptide Self-Assembled Nanoparticles for Efficient Anticancer Drug Delivery
    Authors: B Xu, M Yan, F Zhou, D Cai, W Guo, X Jia, R Liu, T Ma, T Li, F Gao, P Wang, H Lei
    International Journal of Nanomedicine, 2021;16(0):7959-7974.
    Species: Human
    Sample Types: Protein
    Applications: Bioassay
  2. Comparison of Quantification of Target-Specific Accumulation of [18F]F-siPSMA-14 in the HET-CAM Model and in Mice Using PET/MRI
    Authors: J Löffler, C Hamp, E Scheidhaue, D Di Carlo, C Solbach, A Abaei, L Hao, G Glatting, AJ Beer, V Rasche, G Winter
    Cancers, 2021;13(16):.
    Species: Xenografts
    Sample Types: Cell Lysates
    Applications: Western Blot Control
  3. Modelling the internalisation process of prostate cancer cells for PSMA-specific ligands
    Authors: G Winter, A Vogt, LD Jiménez-Fr, A Rinscheid, E Yousefzade, C Solbach, AJ Beer, G Glatting, P Kletting
    Nucl. Med. Biol., 2019;72(0):20-25.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Surface Plasmon Resonance (SPR
  4. Methods for Evaluating Cell-Specific, Cell-Internalizing RNA Aptamers.
    Authors: Hernandez L, Flenker K, Hernandez F, Klingelhutz A, McNamara J, Giangrande P
    Pharmaceuticals (Basel), 0;6(3):295-319.
    Species: Human
    Sample Types: Aptamer
    Applications: CAR-T (Enzyme Assay)
  5. Targeted inhibition of prostate cancer metastases with an RNA aptamer to prostate-specific membrane antigen.
    Authors: Dassie J, Hernandez L, Thomas G, Long M, Rockey W, Howell C, Chen Y, Hernandez F, Liu X, Wilson M, Allen L, Vaena D, Meyerholz D, Giangrande P
    Mol Ther, 0;22(11):1910-22.
    Species: Human
    Sample Types: Aptamer
    Applications: CAR-T (Enzyme Assay)
  6. Structural computational modeling of RNA aptamers.
    Authors: Xu X, Dickey D, Chen S, Giangrande P
    Methods, 0;103(0):175-9.
    Species: Human
    Sample Types: Aptamer
    Applications: Enzyme Assay
  7. Tyramide signal amplification for antibody-overlay lectin microarray: a strategy to improve the sensitivity of targeted glycan profiling.
    Authors: Meany D, Hackler L, Zhang H, Chan D
    J Proteome Res, 0;10(3):1425-31.
    Species: Human
    Sample Types: Protein
    Applications: Glycan Profiling

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Reviews for Recombinant Human PSMA/FOLH1 Protein, CF

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Recombinant Human PSMA/FOLH1 Protein, CF
By Jenna Nguyen on 07/29/2019
Application: Cell migration/motility

Recombinant Human PSMA/FOLH1 Protein, CF
By Anonymous on 05/07/2019
Application: CellProlif