Recombinant Human PSMA/FOLH1 Protein, CF Summary
Product Specifications
Lys44-Ala750, with an N-terminal 6-His tag
Analysis
Product Datasheets
Carrier Free
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.
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.
|
Assay Procedure
- 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
- Dilute rhPSMA to 0.4 µg/mL in Assay Buffer.
- Dilute Substrate to 40 µM with Assay Buffer.
- 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.
- Incubate at 37 °C for 1 hour.
- Stop the reaction by heating at 95 °C for 5 minutes, then cool to room temperature.
- Prepare a 15 mM OPA solution in OPA Buffer.
- Add 250 µL of OPA solution to all vials and vortex.
- Incubate at room temperature for 10 minutes.
- Load 200 µL of reaction and control to plate.
- Read at excitation and emission wavelengths of 330 nm and 450 nm (top read), respectively in endpoint mode.
- 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
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).
- Silver, D.A. et al. (1997) Clin. Cancer Res. 3:81.
- Carter, R.E. et al. (1996) Pro. Natl. Acad. Sci. USA. 93:749.
- Jackson, P.F. and Slusher, B.S. (2001) Curr. Med. Chem. 8:949.
- Heston, W.D. (1997) Urology 49:104.
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.
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Citations: Showing 1 - 8
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Preclinical Characterization of ARX517, a Site-specific Stable PSMA-Targeted Antibody Drug Conjugate for Treatment of Metastatic Castration-Resistant Prostate Cancer
Authors: Skidmore, LK;Mills, D;Kim, JY;Knudsen, NA;Nelson, JD;Pal, M;Wang, J;Gc, K;Gray, MJ;Barkho, W;Nagaraja Shastri, P;Ramprasad, MP;Tian, F;O'Connor, D;Buechler, YJ;Zhang, SS;
Molecular cancer therapeutics
Species: Human
Sample Types:
Applications: Bioassay -
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, 2021-12-03;16(0):7959-7974.
Species: Human
Sample Types: Protein
Applications: Bioassay -
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, 2021-08-09;13(16):.
Species: Xenograft
Sample Types: Cell Lysates
Applications: Western Blot Control -
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., 2019-05-29;72(0):20-25.
Species: Human
Sample Types: Recombinant Protein
Applications: Surface Plasmon Resonance (SPR -
Structural computational modeling of RNA aptamers.
Authors: Xu X, Dickey D, Chen S, Giangrande P
Methods, 2016-03-10;103(0):175-9.
Species: N/A
Sample Types: Aptamer
Applications: Bioassay -
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, 2014-06-23;22(11):1910-22.
Species: Human
Sample Types: Aptamer
Applications: CAR-T (Enzyme Assay) -
Methods for Evaluating Cell-Specific, Cell-Internalizing RNA Aptamers.
Authors: Hernandez L, Flenker K, Hernandez F, Klingelhutz A, McNamara J, Giangrande P
Pharmaceuticals (Basel), 2013-03-14;6(3):295-319.
Species: Human
Sample Types: Aptamer
Applications: CAR-T (Enzyme Assay) -
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, 2011-01-04;10(3):1425-31.
Species: Human
Sample Types: Protein
Applications: Bioassay
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