Recombinant Mouse Complement Factor D Protein, CF

R&D Systems | Catalog # 9207-SE

R&D Systems
100% Guarantee

Key Product Details

  • R&D Systems NS0-derived Recombinant Mouse Complement Factor D Protein (9207-SE)
  • 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

P03953

Applications

Enzyme Activity
View all Complement Factor D/Adipsin Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived mouse Complement Factor D/Adipsin protein
Ile26-Ser259, with a C-terminal 10-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

Ile26 and His35

Predicted Molecular Mass

27 kDa

SDS-PAGE

40-49 kDa, reducing conditions

Activity

Measured by its ability to cleave a colorimetric peptide substrate, N-carbobenzyloxy-Lys-ThioBenzyl ester (Z-Lys-SBzl), in the presence of 5,5’Dithio-bis (2-nitrobenzoic acid) (DTNB). Edwards, K.M. et al. (1999) J. Biol. Chem. 274:30468.
The specific activity is >18 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation, and Storage

9207-SE
Formulation Supplied as a 0.2 μm filtered solution in Tris and NaCl.
Shipping The product is shipped with dry ice or equivalent. 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: Complement Factor D/Adipsin

Complement Factor D, also known as Adipsin, is a serine protease that catalyzes the initial proteolytic step in the alternative pathway of complement. It cleaves the C3b-bound Factor B, resulting in the formation of a C3bBcomplex, which functions as the alternative pathway C3 convertase (1-3). Factor D is primarily expressed in adipose tissue and is released into the circulation as a zymogen with a 5 amino acid (aa) N-terminal activation peptide (4-6). It is activated by MASP-1 mediated proteolytic removal of the activation peptide (6). Factor D triggered complement activation is important for the clearance of cell debris and control of inflammation following tissue damage (7). The C3 convertase generates C3a which binds to C3a R on pancreatic beta cells to induce insulin production and regulate glucose homeostasis (5). Mature mouse Complement Factor D shares 65% and 83% amino acid sequence identity with human and rat Complement Factor D, respectively. Its protease activity is regulated by reversible conformational changes, which is distinct from most serine proteases whose regulation involves either activation by zymogen processing or inactivation by binding to inhibitors. Compared to its cleavage of C3b-bound Factor B, Factor D has much lower activity toward synthetic peptide substrates. However, thioester substrates have been routinely used for assessing Factor D activity (8).

References

  1. Kijlstra, A. and T.T. Berendschot (2015) Ophthalmic Res. 54:64.
  2. Taylor, F.R. et al. (1999) Biochemistry 38:2849.
  3. Ricklin, D. et al. (2010) Nat. Immunol. 11:785.
  4. White, R.T. et al. (1992) J. Biol. Chem. 267:9210.
  5. Lo, J.C. et al. (2014) Cell 158:41.
  6. Takahashi, M. et al. (2010) J. Exp. Med. 207:29.
  7. Cresci, G.A. et al. (2015) Mol. Immunol. 64:9.
  8. Kim, S. et al. (1995) J. Biol. Chem. 270:24399.

Alternate Names

Adipsin, ADN, AMBP-1, CFD, PFD

Entrez Gene IDs

1675 (Human); 11537 (Mouse); 54249 (Rat)

Gene Symbol

CFD

UniProt

P03953

Additional Complement Factor D/Adipsin Products

Product Documents for Recombinant Mouse Complement Factor D 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 Mouse Complement Factor D Protein, CF

For research use only

Related Research Areas

Complement Pathway

Customer Reviews for Recombinant Mouse Complement Factor D Protein, CF

There are currently no reviews for this product. Be the first to review Recombinant Mouse Complement Factor D Protein, CF and earn rewards!

Have you used Recombinant Mouse Complement Factor D 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 Mouse Complement Factor D Protein, CF (9207-SE):

Materials
  • Assay Buffer: 50 mM Tris, 1 M NaCl, pH 7.5
  • Recombinant Mouse Complement Factor D/Adipsin (rmFactor D) (Catalog # 9207-SE)
  • Substrate: Z-Lys-SBzl (Bachem, Catalog # M-1300), 10 mM stock in DMSO
  • 5',5'-Dithiobis(2-nitrobenzoic acid) (DTNB) (Sigma, Catalog # D8130), 10 mM stock in DMSO
  • 96-well Clear Plate (Catalog # DY990)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Dilute rmFactor D to 20 µg/mL in Assay Buffer.
  2. Dilute substrate to 400 µM in Assay Buffer containing 400 µM DTNB.
  3. Load 50 µL of 20 µg/mL rmFactor D to plate, and start the reaction by adding 50 µL of substrate/DTNB mixture.  Include a Substrate Blank containing 50 µL of Assay Buffer and 50 µL of substrate/DTNB mixture.
  4. Read in kinetic mode for 20 minutes at an absorbance of 405 nm.

     Specific Activity (pmol/min/µg) =

Adjusted Vmax* (OD/min) x well volume (L) x 1012 pmol/mol
ext. coeff** (M-1cm-1) x path corr.*** (cm) x amount of enzyme (µg)


     *Adjusted for Substrate Blank
    **Using the extinction coefficient 13260 M-1cm-1
    ***Using the path correction 0.32 cm
    Note: the output of many spectrophotometers is in mOD.

Per Well:

  • rmFactor D: 1.0 µg
  • Substrate: 200 µM
  • DTNB: 200 µM

FAQs

No product specific FAQs exist for this product.

View all FAQs for Proteins and Enzymes