Recombinant Human Complement Factor D Protein, CF

R&D Systems | Catalog # 1824-SE

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Key Product Details

  • R&D Systems NS0-derived Recombinant Human Complement Factor D Protein (1824-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

P00746

Applications

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

Product Specifications

Source

Mouse myeloma cell line, NS0-derived human Complement Factor D/Adipsin protein
Ile26-Ala253, 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 & His35

Predicted Molecular Mass

26 kDa & 25 kDa

SDS-PAGE

27 kDa & 26 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 >70 pmol/min/µg, as measured under the described conditions.

Reviewed Applications

Read 1 review rated 5 using 1824-SE in the following applications:

Formulation, Preparation, and Storage

1824-SE
Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl, CaCl2 and Glycerol.
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.

Background: Complement Factor D/Adipsin

Complement Factor D is a serine protease that catalyzes the initial proteolytic step in the alternative pathway of complement. Expressed in adipose tissue at high levels, factor D is also known as adipsin (1). It is an exceptionally specific protease and the only known protein substrate is factor B in complex with C3 (2). Factor D protease activity is regulated by reversible conformational changes, which differs from the majority of serine proteases whose regulation involves either activation by processing of the zymogens or inactivation by binding of the inhibitors. Compared to its physiologically important proteolytic activity, factor D has much lower activity toward synthetic peptide substrates. However, thioester substrates have been routinely used for assessing factor D activity (3).

References

  1. White, R.T. et al. (1992) J. Biol. Chem. 267:9210.
  2. Taylor, F.R. et al. (1999) Biochemistry 38:2849.
  3. 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

P00746

Additional Complement Factor D/Adipsin Products

Product Documents for Recombinant Human Complement Factor D Protein, CF

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Recombinant Human Complement Factor D Protein, CF

For research use only

Related Research Areas

Complement Pathway

Citations for Recombinant Human Complement Factor D Protein, CF

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    Application: Apoptosis assay
    Verified Customer | Posted 07/27/2018

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Protocols

View specific protocols for Recombinant Human Complement Factor D Protein, CF (1824-SE):

Materials
  • Assay Buffer: 50 mM Tris, 1 M NaCl, pH 7.5
  • Recombinant Human Complement Factor D/Adipsin (rhFactor D) (Catalog # 1824-SE)
  • Substrate: Z-Lys-SBzl (Bachem, Catalog # M-1300), 10 mM stock in DMSO
  • 5,5’Dithio-bis-(2-nitrobenzoic acid) (DTNB) (Sigma, Catalog # D-8130), 10 mM stock in DMSO
  • 96 Well Clear Plate (Costar, Catalog # 92592)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Dilute rhFactor D to 5 ng/µL in Assay Buffer.
  2. Dilute Substrate to 200 µM in Assay Buffer with 200 µM DTNB.
  3. Load 50 µL of the diluted rhFactor D into a clear plate, and start the reaction by adding 50 µL of Substrate/DTNB mixture to wells. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL Substrate mixture without any rhFactor D.
  4. Read in kinetic mode for 20 minutes at an absorbance of 405 nm.
  5. Calculate specific activity:

     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 0.320 cm
     Note: the output of many spectrophotometers is in mOD Per Well:
  • rhFactor D: 0.25 µg
  • DTNB: 100 µM
  • Substrate: 100 µM

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