Human Complement Factor D Quantikine ELISA Kit

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Human Complement Factor D/Adipsin ELISA Standard Curve
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Human Complement Factor D Quantikine ELISA Kit Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
3.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 uL), Serum (10 uL), EDTA Plasma (10 uL), Heparin Plasma (10 uL), Urine (50 uL)
Sensitivity
0.025 ng/mL
Assay Range
0.3 - 20 ng/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Urine)
Specificity
Natural and recombinant human Complement Factor D
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
Interference
No significant interference observed with available related molecules.

Product Summary

The Quantikine Human Complement Factor D Immunoassay is a 3.5 hour sandwich ELISA designed to measure human Complement Factor D in cell culture supernates, serum, plasma, and urine. It contains NS0-expressed recombinant human Complement Factor D and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human Complement Factor D showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring human Complement Factor D.

Precision

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision

Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Urine

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean (pg/mL) 0.86 3.12 8.13 0.831 3.04 7.73
Standard Deviation 0.047 0.183 0.552 0.074 0.211 0.506
CV% 6.4 5.9 5.8 9 6.9 6.6

Recovery

The recovery of Complement Factor D spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 104 97-117
Urine (n=4) 100 87-115

Linearity

To assess the linearity of the assay, samples containing and/or spiked with high concentrations of Complement Factor D were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human Complement Factor D/Adipsin ELISA Linearity

Scientific Data

Human Complement Factor D/Adipsin ELISA Standard Curve

Product Datasheets

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Preparation and Storage

Shipping
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

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 is an exceptionally specific protease and the only known protein substrate is factor B in complex with C3.3 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.

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
&#9888; WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

Assay Procedure

Refer to the product for complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

  3. 50 µL Assay Diluent
  4.   Add 50 µL of Assay Diluent to each well.

  5. 50 µL Standard, Control, or Sample
  6.   Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
  7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

  8. 100 µL Conjugate
  9.   Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 1 hour.
  10.   Aspirate and wash 4 times.

  11. 100 µL Substrate Solution
  12.   Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.

  13. 100 µL Stop Solution
  14.   Add 100 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

Citations for Human Complement Factor D Quantikine ELISA Kit

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.

4 Citations: Showing 1 - 4
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  1. Adipsin Serum Concentrations and Adipose Tissue Expression in People with Obesity and Type 2 Diabetes
    Authors: M Milek, Y Moulla, M Kern, C Stroh, A Dietrich, MR Schön, D Gärtner, T Lohmann, M Dressler, P Kovacs, M Stumvoll, M Blüher, E Guiu-Jurad
    International Journal of Molecular Sciences, 2022-02-17;23(4):.
    Species: Human
    Sample Types: Serum
  2. Danicopan: an oral complement factor D inhibitor for paroxysmal nocturnal hemoglobinuria
    Authors: AM Risitano, AG Kulasekara, JW Lee, JP Maciejewsk, R Notaro, R Brodsky, M Huang, M Geffner, P Browett
    Haematologica, 2021-12-01;0(0):0.
    Species: Human
    Sample Types: Serum
  3. Small-molecule Factor D inhibitors selectively block the alternative pathway of complement in paroxysmal nocturnal hemoglobinuria and atypical hemolytic uremic syndrome
    Haematologica, 2016-11-03;0(0):.
    Species: Human
    Sample Types: Serum
  4. Plasma complement components and activation fragments: associations with age-related macular degeneration genotypes and phenotypes.
    Authors: Reynolds R, Hartnett ME, Atkinson JP, Giclas PC, Rosner B, Seddon JM
    Invest. Ophthalmol. Vis. Sci., 2009-08-06;50(12):5818-27.
    Species: Human
    Sample Types: Plasma

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