Overview

  • Product name

    Human Complement Factor D ELISA Kit
    See all Factor D/CFD kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    serum 8 2.4%
    Inter-assay
    Sample n Mean SD CV%
    serum 3 4.3%
  • Sample type

    Cell culture supernatant, Urine, Serum, Hep Plasma, EDTA Plasma, Cit plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    2.6 pg/ml
  • Range

    15.63 pg/ml - 1000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell culture supernatant 96 87% - 102%
    Urine 88 83% - 90%
    Serum 98 92% - 105%
    Hep Plasma 95 82% - 102%
    EDTA Plasma 93 84% - 102%
    Cit plasma 94 90% - 100%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Human
  • Product overview

    Complement Factor D in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Complement Factor D protein in human serum, plasma, urine, and cell culture supernatant.


     


    The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB Development Solution is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.


    Complement Factor D, also known as adipsin, is a serine protease that is indispensable for the initiation of complement activation by the alternative pathway. Upon activation through reversible substrate-induced conformational change into an active enzyme, factor D functions to cleave the C3b-bound factor B, resulting in the formation of C3bBb complex, which is the alternative pathway C3 convertase. Human complement factor D is synthesized as a 253 amino acid (aa) precursor that contains a signal peptide (aa 1-20), a five residue activation-/pro-peptide (aa 21-25), and the mature chain (aa 26-253). Under physiological conditions, mature factor D lacking the activation peptide circulates as an inactive enzyme and requires interaction with its natural substrate, C3b-bound factor B, for activation of its catalytic activity (2). Mature human factor D shares 98%, 96%, 84%, and 66% aa sequence homology with the chimpanzee, rhesus monkey, porcine, and mouse protein, respectively.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Pre-coated microplate (12 x 8 well strips)

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Human Complement Factor D Capture Antibody 1 x 600µl
    10X Human Complement Factor D Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    Antibody Diluent 4BI 1 x 6ml
    Human Complement Factor D Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS (ab193972) 1 x 50ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • Research areas

  • Function

    Factor D cleaves factor B when the latter is complexed with factor C3b, activating the C3bbb complex, which then becomes the C3 convertase of the alternate pathway. Its function is homologous to that of C1s in the classical pathway.
  • Involvement in disease

    Defects in CFD are the cause of complement factor D deficiency (CFD deficiency) [MIM:134350]. CFD deficiency predisposes to invasive meningococcal disease.
  • Sequence similarities

    Belongs to the peptidase S1 family.
    Contains 1 peptidase S1 domain.
  • Cellular localization

    Secreted.
  • Information by UniProt
  • Alternative names

    • Adipsin
    • Adipsin/complement factor D
    • Adn
    • C3 convertase activator
    • CFAD_HUMAN
    • CFD
    • Complement factor D
    • complement factor D preproprotein
    • D component of complement
    • DF
    • FactorD
    • PFD
    • Properdin factor D
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab243686 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Sandwich ELISA Use at an assay dependent concentration.

Images

  • SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

     

  • Standard Curve comparison between human Complement Factor D SimpleStep ELISA kit and traditional ELISA kit from leading competitor. SimpleStep ELISA kit shows increased sensitivity.

  • Example of human Complement Factor D standard curve in Sample Diluent NS.

  • The concentrations of Complement Factor D were measured in duplicates, interpolated from the Complement Factor D standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 1:1,000, plasma (citrate) 1:1,000, plasma (EDTA) 1:1,000and plasma (heparin) 1:1,000. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Complement Factor D concentration was determined to be 944.1 ng/mL in serum, 721.8 ng/mL in plasma (citrate), 860.5 ng/mL in plasma (EDTA), and 321.7 ng/mL in plasma (heparin).

  • The concentrations of Complement Factor D were measured in duplicates, interpolated from the Complement Factor D standard curves and corrected for sample dilution. Undiluted samples are as follows: male urine 100%, female urine 50%, and U937 supernatant 12.5%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Complement Factor D concentration was determined to be 958.8 pg/mL in male urine, 494.7 pg/mL in female urine and 4,734.2 pg/mL in U937 supernatant.

     

  • Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Complement Factor D concentration was determined to be 975.9 ng/mL with a range of 556.8 – 1,603.5 ng/mL.

     

     

  • Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Complement Factor D concentration was determined to be 1,142.5 ng/mL with a range of 867.1 – 1,958.8 ng/mL.

     

     

Protocols

References

ab243686 has not yet been referenced specifically in any publications.

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