Recombinant Human Serpin A3 Protein, CF

R&D Systems | Catalog # 1295-PI

R&D Systems
100% Guarantee

Key Product Details

  • R&D Systems NS0-derived Recombinant Human Serpin A3 Protein (1295-PI)
  • 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

NS0

Accession Number

P01011

Applications

Inhibition Activity
View all Serpin A3/alpha 1-Antichymotrypsin Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived human Serpin A3/alpha 1-Antichymotrypsin protein
Asn26-Ala423, with a C-terminal 10-His tag

Purity

>90%, by SDS-PAGE under reducing conditions and visualized by silver stain.

Endotoxin Level

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

N-terminal Sequence Analysis

Asn26

Predicted Molecular Mass

46 kDa

SDS-PAGE

60-65 kDa, reducing conditions

Activity

Measured by its ability to inhibit chymotrypsin cleavage of a fluorogenic peptide substrate MCA-RPKPVE-Nval-WRK(Dnp)-NH2 (Catalog # ES002).
The IC50 value is <8 nM, as measured under the described conditions.

Formulation, Preparation, and Storage

1295-PI
Formulation Lyophilized from a 0.2 μm filtered solution in HEPES and NaCl.
Reconstitution Reconstitute at 100 μg/mL in sterile 25 mM HEPES and 150 mM NaCl, pH 7.8.
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 reconstitution.

Calculators

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.

=
÷

Background: Serpin A3/alpha 1-Antichymotrypsin

Serpin A3 is a member of the Serpin superfamily of the serine protease inhibitors (1). It is synthesized primarily in the liver and secreted as one of the most abundant serpins in plasma (2). It is known to inhibit several serine proteases including chymotrypsin, cathepsin G, chymase, kallikrein 3/prostate specific antigen, and unidentified ectoenzymes that process pro-macrophage stimulating protein (1‑5). Serpin A3 is a major constituent of the plaques associated with Alzheimer’s disease and an inhibitor of amyloid beta peptide degradation (1‑6). Deficiency in Serpin A3 activity due to a point mutation (Leu55Pro) is associated with chronic obstructive pulmonary disease (7). Human Serpin A3 is synthesized as a 423 amino acid precursor (8, 9). The mature protein is secreted and has two forms that differ in their N‑termini (10). One of the forms starting at Asn26 was expressed and purified.

References

  1. Silverman, G.A. et al. (2001) J. Biol. Chem. 276:33293.
  2. Kalsheker, N. et al. (2002) Biochem. Soc. Trans. 30:93.
  3. Duranto, J. et al. (1998) Biochemistry 37:11239.
  4. Hsieh, M.-C. and B.S. Cooperman (2002) Biochemistry 41:2990.
  5. Skeel, A. and E.J. Leonard (2001) J. Biol. Chem. 276:21932.
  6. Abraham, C.R. et al. (2000) Ann. N. Y. Acad. Sci. 920:245.
  7. Gooptu, B. et al. (2000) Proc. Natl. Acad. Sci. USA 97:67.
  8. Chandra, T. et al. (1983) Biochemistry 22:5055.
  9. Morii, M. and J. Travis (1983) J. Biol. Chem. 258:12749.
  10. Lindmark, B. et al. (1989) Biochim. Biophys. Acta 997:90.

Alternate Names

alpha 1-Antichymotrypsin

Entrez Gene IDs

12 (Human)

Gene Symbol

SERPINA3

UniProt

P01011

Additional Serpin A3/alpha 1-Antichymotrypsin Products

Product Documents for Recombinant Human Serpin A3 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.

Download SDS

Product Specific Notices for Recombinant Human Serpin A3 Protein, CF

For research use only

Citations for Recombinant Human Serpin A3 Protein, CF

Customer Reviews for Recombinant Human Serpin A3 Protein, CF

There are currently no reviews for this product. Be the first to review Recombinant Human Serpin A3 Protein, CF and earn rewards!

Have you used Recombinant Human Serpin A3 Protein, CF?

Submit a review and receive an Amazon gift card!

$25/€18/£15/$25CAN/¥2500 Yen for a review with an image

$10/€7/£6/$10CAN/¥1110 Yen for a review without an image

Submit a review
Amazon Gift Card

Protocols

View specific protocols for Recombinant Human Serpin A3 Protein, CF (1295-PI):

Materials
  • Assay Buffer: 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, 0.05% (w/v) Brij-35, pH 7.5 (TCNB)
  • Recombinant Human Serpin A3/ alpha 1-Antichymotrypsin (rhSerpin A3) (Catalog # 1295-PI)
  • Chymotrypsin (Sigma, Catalog # 3142), 40-80 units/mg, prepare a 1 mg/mL stock in 1 mM HCl
  • Substrate: MCA-Arg-Pro-Lys-Pro-Val-Glu-NVAL-Trp-Arg-Lys(DNP)-NH2 (Catalog # ES002), 2 mM stock in DMSO
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent

  1. Dilute Chymotrypsin to 0.6 µg/mL in Assay Buffer.
  2. Prepare a curve of rhSerpin A3 (MW: 46,373 kDa) in Assay Buffer. Make the following serial dilutions including the most concentrated dilution possible: (most concentrated), 1000, 500, 250, 175, 120, 80, 40, 20, and 5 nM.
  3. Mix equal volumes of rhSerpin A3 curve dilutions and 0.6 µg/mL Chymotrypsin in microtubes. Include two enzyme controls of equal volumes of Assay Buffer and 0.6 µg/mL Chymotrypsin.
  4. Incubate reaction mixtures at 37 °C for 30 minutes.
  5. Dilute incubated reaction mixtures by five fold in Assay Buffer.
  6. Dilute Substrate to 20 µM in Assay Buffer.
  7. In a plate load 50 µL of the diluted reaction mixtures to wells, and start the reaction by adding 50 µL of 20 µM Substrate.
  8. Read at excitation and emission wavelengths of 320 nm and 405 nm (top read), respectively, in kinetic mode for 5 minutes.
  9. Derive the 50% inhibiting concentration (IC50) for rhSerpin A3 by ploting RFU/min (or specific activity) vs concentration with 4-PL fitting.
  10. The specific activity for Chymotrypsin at each point may be determined using the following formula (if needed):

     Specific Activity (pmol/min/µg) =

 Adjusted Vmax* (RFU/min) x Conversion Factor** (pmol/RFU)
amount of enzyme (µg)

     *Adjusted for Substrate Blank

     **Derived using calibration standard MCA-Pro-Leu-OH (Bachem, Catalog # M-1975).

Per Well:

  • Chymotrypsin: 0.003 µg
  • rhSerpin A3: (most concentrated), 50, 25, 12.5, 8.75, 6, 4, 2, 1 and 0.25 nM
  • Substrate: 10 µM

FAQs

No product specific FAQs exist for this product.

View all FAQs for Proteins and Enzymes