Recombinant Human Complement Factor D Protein, CF

Name: Recombinant Human Complement Factor D Protein, CF
Brand: R&D Systems
SKU: 1824-SE
Rating: 5 (1 reviews)

Catalog #: 1824-SE
2 Citations 1 Review Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
1824-SE-010

R&D Systems Recombinant Proteins and Enzymes

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All Complement Factor D/Adipsin Proteins and Enzymes

Product Details

Citations (2)

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Reviews (1)

Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Human Complement Factor D Protein, CF Summary

Product Specifications

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.

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.

Source

Mouse myeloma cell line, NS0-derived human Complement Factor D/Adipsin protein
Ile26-Ala253, with a C-terminal 10-His tag

Accession #

P00746

N-terminal Sequence
Analysis

Ile26 & His35

Predicted Molecular Mass

26 kDa & 25 kDa

SDS-PAGE

27 kDa & 26 kDa, reducing conditions

Product Datasheets

1824-SE

Product Datasheet

COA

SDS

Carrier Free

What does CF mean?

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.

What formulation is right for me?

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.

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.

Assay Procedure

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

Dilute rhFactor D to 5 ng/µL in Assay Buffer.
Dilute Substrate to 200 µM in Assay Buffer with 200 µM DTNB.
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.
Read in kinetic mode for 20 minutes at an absorbance of 405 nm.
Calculate specific activity:

Specific Activity (pmol/min/µg) =	Adjusted V_max* (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

Reconstitution Calculator

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

White, R.T. et al. (1992) J. Biol. Chem. 267:9210.
Taylor, F.R. et al. (1999) Biochemistry 38:2849.
Kim, S. et al. (1995) J. Biol. Chem. 270:24399.

Entrez Gene IDs

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

Alternate Names

Adipsin; ADN; ADNcomplement factor D; AMBP-1; C3 convertase activator; CFD; complement factor D (adipsin); complement factor D preproprotein; Complement Factor D; D component of complement (adipsin); DF; EC 3.4.21; EC 3.4.21.46; PFD; Properdin factor DADIPSIN

Related Research Areas

Citations for Recombinant Human Complement Factor D 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.

2 Citations: Showing 1 - 2
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Adiponectin inhibits insulin function in primary trophoblasts by PPARalpha-mediated ceramide synthesis.
Authors: Aye I, Gao X, Weintraub S, Jansson T, Powell T
Mol Endocrinol, 2014-02-25;28(4):512-24.
Species: Human
Sample Types: Whole Cells
Applications: Bioassay
A multiplex immunoassay for human adipokine profiling.
Authors: Schipper HS, De Jager W, van Dijk ME, Meerding J, Zelissen PM, Adan RA, Prakken BJ, Kalkhoven E
Clin. Chem., 2010-06-08;56(0):1320.
Applications: ELISA (Standard)