Human Plasminogen Protein, CF Summary
Product Specifications
The human plasma used for the isolation of this product was certified by the supplier to be HIV-1 and HBsAg negative at the time of shipment. Human blood products should always be treated in accordance with universal handling precautions.
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.
1939-SE
| Formulation | Lyophilized from a 0.2 μm filtered solution in Tris and NaCl. |
| Reconstitution | Reconstitute at 1 mg/mL in sterile 25 mM Tris and 150 mM NaCl, pH 8.0. |
| Shipping | The product is shipped at ambient temperature. 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
- Activation Buffer: 50 mM Tris, 0.01% Tween® 20, pH 8.5
- Assay Buffer: 0.1 M Tris, 0.1 M NaCl, pH 7.5
- Human Plasminogen (hPLG) (Catalog # 1939-SE)
- Recombinant Human u-Plasminogen Activator (uPA)/Urokinase (rhuPA) (Catalog # 1310-SE)
- Substrate: SUC-Ala-Phe-Lys-AMC (Bachem, Catalog # I-1330 ), 10 mM stock in DMSO
- F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
- Fluorescent Plate Reader (Model: Spectramax Gemini EM by Molecular Devices) or equivalent
- Dilute hPLG to 200 µg/mL in Activation Buffer.
- Dilute rhuPA to 4 µg/mL in Activation Buffer.
- Combine 25 µL of diluted hPLG with 25 µL of diluted rhuPA for final concentrations of 100 µg/mL and 2 µg/mL respectively.
- Incubate at 37 °C for 15 minutes.
- Dilute activated hPLG to 2 ng/µL in Assay Buffer.
- Dilute Substrate to 200 µM in Assay Buffer.
- Load 50 mL of 2 ng/µL hPLG in a black well plate, and start the reaction by adding 50 µL of 200 µM substrate. Include a Substrate Blank containing 50 µL of Assay Buffer and 50 µL of 200 µM Substrate.
- Read at excitation and emission wavelengths of 380 nm and 460 nm (top read), respectively in kinetic mode for 5 minutes.
- Calculate specific activity:
| Specific Activity (pmol/min/µg) = |
Adjusted Vmax* (RFU/min) x Conversion Factor** (pmol/RFU) |
| amount of enzyme (µg) |
**Derived using calibration standard 7-Amino, 4-Methyl Coumarin (AMC) (Sigma, Catalog # A-9891). Per Well:
- hPLG: 0.1 µg
- Substrate: 100 µM
Reconstitution Calculator
Background: Plasminogen
Plasminogen (PLG) is the precursor of plasmin, an active serine protease that dissolves the fibrin of blood clots and acts in many other processes such as embryonic development, tissue remodeling, inflammation and tumor invasion (1, 2). Synthesized in the kidney, PLG is found in plasma and many extracellular fluids. Activated by u- or t-plasminogen activator, the single-chain PLG (amino acid residues 20‑810) is converted to plasmin, which consists of disulfide bond-linked heavy chain A (residues 20-580) and light chain B (residues 581‑810). Heavy chain A contains 5 kringle domains and light chain B corresponds to the serine protease domain. A fragment consisting of the first 4 kringle domains has been named as angiostatin, a novel angiogenesis inhibitor (3, 4).
- Petersen, T.E. et al. (1990) J. Biol. Chem. 265:6104.
- Forsgren, M. et al. (1987) FEBS Lett. 213:254.
- O’Reilly, M.S. et al. (1994) Cell 79:315.
- Sim, B.K. et al. (1997) Cancer Res. 57:1329.
Citations for Human Plasminogen 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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Human plasma plasminogen internalization route in Plasmodium falciparum-infected erythrocytes
Authors: S El Chamy M, MY Icimoto, PMS Melo, A Budu, R Coimbra, ML Gazarini, AK Carmona
Malar. J., 2020-08-26;19(1):302.
Species: Complex Species Category
Sample Types: Whole Cells
Applications: Bioassay -
Plasminogen promotes cholesterol efflux by the ABCA1 pathway
Authors: N Pamir, PM Hutchins, GE Ronsein, H Wei, C Tang, R Das, T Vaisar, E Plow, V Schuster, CA Reardon, R Weinberg, DA Dichek, S Marcovina, GS Getz, JW Heinecke
JCI Insight, 2017-08-03;2(15):.
Species: Human
Sample Types: Whole Cells
Applications: Bioassay -
A novel role for plasmin-mediated degradation of opsonizing antibody in the evasion of host immunity by virulent, but not attenuated, Francisella tularensis.
Authors: Crane DD, Warner SL, Bosio CM
J. Immunol., 2009-09-14;183(7):4593-600.
Species: Bacteria - Francisella tularensis
Sample Types: Buffer
Applications: ELISA Developmet
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