Recombinant Human Complement Factor D (carrier-free)

Pricing & Availability
Regulatory Status
RUO
Other Names
DF, ADN, PFD, Adipsin, Properdin Factor D, C3 Convertase Activator, Complement Factor D (Adipsin)
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Human_Complement-Factor-D_RECOM_CF_1_051820
Recombinant human complement factor D activity is 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). The specific activity is > 70 pmol/min/µg.
  • Human_Complement-Factor-D_RECOM_CF_1_051820
    Recombinant human complement factor D activity is 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). The specific activity is > 70 pmol/min/µg.
  • Human_Complement-Factor-D_RECOM_CF_2_051820
    Stability Testing for Recombinant Human Complement Factor D. Recombinant human complement factor D was aliquoted in PBS pH 7.2. One aliquot was frozen and thawed four times (4x Freeze/Thaw) and compared to the control that was kept at 4°C (Control). The samples were tested for their 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). The specific activity is > 70 pmol/min/µg.
Cat # Size Price Quantity Check Availability Save
791808 500 µg 1575,00 CHF
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791802 10 µg 185,00 CHF
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791804 25 µg 310,00 CHF
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791806 100 µg 650,00 CHF
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Description

Complement Factor D (Adipsin) is a serine protease with restricted substrate specificity. It cleaves a single lysine-arginine bond in complement factor B, only when B is complexed with complement component C3 (C3b-bound factor B). Factor D limits the rate of the alternative complement pathway. Adipose tissue is the main source of factor D. Also, monocytes and macrophages express this protein. Studies in Masp1/3−/− mice (Mannose-binding lectin-associated serine protease) identified that factor D is produced as a pro-factor D with the activation peptide QPRGR at its N terminus. Masp1/3−/− mice do not show alternative complement pathway activation in serum; suggesting that Masp1/3−/− mice do not change pro-factor D to its active form. Adipocytes produce complement component 3, and complement factors B and D. Factor D induces adipocyte differentiation through the induction of C3a and C3aR, thus the interaction between C3a and C3aR induces triglyceride synthesis. Studies with Adipsin -/- mice suggest that factor D improves β cell function in diabetes. Factor D is associated to the onset of pulmonary arterial hypertension in systemic sclerosis. In addition, factor D seems to be a prognostic biomarker in patients with coronary artery diseases.

Product Details
Technical Data Sheet (pdf)

Product Details

Source
Human Complement Factor D, amino acid Ile26-Ala253 (Accession #: P00746) was expressed in CHO cells. The C-terminus contains 8His- GGQ tag.
Molecular Mass
The 241 amino acid recombinant protein has a predicted molecular mass of approximately 25.9 kD. The DTT-reduced and non reduced proteins migrate at approximately 28 kD by SDS-PAGE. The predicted N-terminal amino acid is Ile.
Purity
> 95%, as determined by Coomassie stained SDS-PAGE
Formulation
0.22 µm filtered protein solution is in PBS, pH 7.2
Endotoxin Level
Less than 0.1 EU per µg cytokine as determined by the LAL method
Concentration
10 and 25 µg sizes are bottled at 200 µg/mL. 100 µg and larger sizes are lot-specific and bottled at the concentration indicated on the vial (please contact technical support for concentration, or use our Lookup tool if you have a lot number.)
Storage & Handling
Unopened vial can be stored at -20°C or -70°C for six months. For maximum results, quick spin vial prior to opening. Avoid repeated freeze/thaw cycles.
Activity
Complement factor D cleaves a colorimetric peptide substrate, N-carbobenzyloxy-Lys-Thiobenzyl ester (Z-Lys-SBzl), in the presence of 5,5’Dithio-bis (2-nitrobenzoic acid) (DTNB). The specific activity is > 70 pmol/min/µg, as measured under the described conditions.
Application

Bioassay

Application Notes

Assay Procedure

Recombinant Human complement Factor D/Adipsin activity is 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). The increase of the product is monitored by increase in intensity of absorbance at 405 nm.

Materials

  • Assay buffer: 50 mM Tris, 1 M NaCl, pH 7.5
  • Recombinant human Complement Factor D/Adipsin
  • Substrate: Z-Lys-SBzl (Bachem, Cat. No. M-1300), 10 mM stock in DMSO
  • 5,5’Dithio-bis-(2-nitrobenzoic acid) (DTNB) (Sigma, Cat. No. D-8130), 10 mM stock in DMSO
  • 96-well clear plate (Costar, Cat. No. 92592)
  • Plate reader (Model: SpectraMax Plus by Molecular Devices) or equivalent


Procedure

  1. Dilute the recombinant factor D to 5 ng/µL in assay buffer.
  2. Dilute the substrate to 200 µM in assay buffer with 200 µM DTNB.
  3. Load 50 µL of the diluted recombinant factor D into a clear plate, and start the reaction by adding 50 µL of substrate/DTNB mixture to the wells. Include a substrate blank containing 50 µL assay buffer and 50 µL substrate mixture without any recombinant Factor D.
  4. Read in kinetic mode for 5 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

  • Recombinant factor D: 0.25 µg
  • DTNB: 100 µM
  • Substrate: 100 µM


BioLegend carrier-free recombinant proteins provided in liquid format are shipped on blue-ice. Our comparison testing data indicates that when handled and stored as recommended, the liquid format has equal or better stability and shelf-life compared to commercially available lyophilized proteins after reconstitution. Our liquid proteins are verified in-house to maintain activity after shipping on blue ice and are backed by our 100% satisfaction guarantee. If you have any concerns, contact us at tech@biolegend.com.

Antigen Details

Structure
Monomer
Distribution

Adipose tissue, monocytes, macrophages, microvascular endothelial cells

Function
Alternative pathway of the complement system, cleaves complement factor B, promotes adipocyte differentiation
Ligand/Receptor
Complement factor B
Bioactivity
Complement factor D cleaves a colorimetric peptide substrate, N-carbobenzyloxy-Lys-Thiobenzyl ester (Z-Lys-SBzl), in the presence of 5,5’Dithio-bis (2-nitrobenzoic acid) (DTNB).
Cell Type
Endothelial cells, Macrophages, Monocytes
Biology Area
Cardiovascular Biology, Cell Proliferation and Viability, Complement, Immunology, Innate Immunity
Molecular Family
Enzymes and Regulators, Innate Immune Signaling, Proteases
Antigen References
  1. White RT, et al. 1992. J Biol Chem. 267:9210.
  2. Pattrick M, et al. 2009. Mol Immunol. 46:755.
  3. Mamane Y, et al. 2009. Diabetes. 58:2006.
  4. Lo JC, et al. 2014. Cell. 158:41.
  5. Song NJ, et al. 2016. PLoS One. 11:e0162228.
  6. Korman BD, et al. 2017. Arthritis Rheumatol. 69:2062.
  7. Ohtsuki T, et al. 2019. J Am Heart Assoc. 8:e013716.
Gene ID
1675 View all products for this Gene ID
UniProt
View information about Complement Factor D on UniProt.org

Related FAQs

Why choose BioLegend recombinant proteins?

     • Each lot of product is quality-tested for bioactivity as indicated on the data sheet.
     • Greater than 95% Purity or higher, tested on every lot of product.
     • 100% Satisfaction Guarantee for quality performance, stability, and consistency.
     • Ready-to-use liquid format saves time and reduces challenges associated with reconstitution.
     • Bulk and customization available. Contact us.
     • Learn more about our Recombinant Proteins.

How does the activity of your recombinant proteins compare to competitors?

We quality control each and every lot of recombinant protein. Not only do we check its bioactivity, but we also compare it against other commercially available recombinant proteins. We make sure each recombinant protein’s activity is at least as good as or better than the competition’s. In order to provide you with the best possible product, we ensure that our testing process is rigorous and thorough. If you’re curious and eager to make the switch to BioLegend recombinants, contact your sales representative today!

What is the specific activity or ED50 of my recombinant protein?

The specific activity range of the protein is indicated on the product datasheets. Because the exact activity values on a per unit basis can largely fluctuate depending on a number of factors, including the nature of the assay, cell density, age of cells/passage number, culture media used, and end user technique, the specific activity is best defined as a range and we guarantee the specific activity of all our lots will be within the range indicated on the datasheet. Please note this only applies to recombinants labeled for use in bioassays. ELISA standard recombinant proteins are not recommended for bioassay usage as they are not tested for these applications.

Have your recombinants been tested for stability?

Our testing shows that the recombinant proteins are able to withstand room temperature for a week without losing activity. In addition the recombinant proteins were also found to withstand four cycles of freeze and thaw without losing activity.

Does specific activity of a recombinant protein vary between lots?

Specific activity will vary for each lot and for the type of experiment that is done to validate it, but all passed lots will have activity within the established ED50 range for the product and we guarantee that our products will have lot-to-lot consistency. Please conduct an experiment-specific validation to find the optimal ED50 for your system.

How do you convert activity as an ED50 in ng/ml to a specific activity in Units/mg?
Use formula Specific activity (Units/mg) = 10e6/ ED50 (ng/mL)
Go To Top Version: 1    Revision Date: 05.18.2020

For research use only. Not for diagnostic use. Not for resale. BioLegend will not be held responsible for patent infringement or other violations that may occur with the use of our products.

 

*These products may be covered by one or more Limited Use Label Licenses (see the BioLegend Catalog or our website, www.biolegend.com/ordering#license). BioLegend products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products, reverse engineer functionally similar materials, or to provide a service to third parties without written approval of BioLegend. By use of these products you accept the terms and conditions of all applicable Limited Use Label Licenses. Unless otherwise indicated, these products are for research use only and are not intended for human or animal diagnostic, therapeutic or commercial use.

 

BioLegend Inc., 8999 BioLegend Way, San Diego, CA 92121 www.biolegend.com
Toll-Free Phone: 1-877-Bio-Legend (246-5343) Phone: (858) 768-5800 Fax: (877) 455-9587

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