Complement C4d assay

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The Complement C4d assay is an enzyme-linked immunosorbent assay (ELISA) for the semi-quantitative determination of C4d in human plasma.

The complement system plays an essential role in autoimmune and infectious diseases. There are three pathways of complement activation; the classical, the alternative and the lectin pathway. The C4d protein is a product of the classical and the lectin pathways. 

The Svar Life Science Complement C4d assay is a highly specific C4d Elisa kit for measurement of C4d in human plasma. 

C4d can be used as a complement activity marker for the classical and lectin pathways. 

The assay is targeting a unique cleavage epitope on C4d, which means that only the C4d is measured.

 

Product code
COMPL C4d RUO
Format
ELISA
Tests
Break apart microtitration strips (12x8) 96 wells
Calculation
Semi-Quantitative
Incubation time
60+30+30 min
Detection system
450 nm
Availability
For Research Use Only. Not for use in diagnostic procedures.

The Complement C4d assay is an enzyme-linked immunosorbent assay (ELISA) for the semi-quantitative determination of C4d in human plasma.

The product is intended for professional use. The result shall not be used for clinical diagnosis or patient management. FOR RESEARCH USE ONLY.

The device is a colorimetric sandwich ELISA. Samples are diluted in assay diluent and 100µL diluted sample is transferred to the microtiter wells and incubated at room temperature for 60 minutes. During this first incubation C4d in the sample is captured by the anti-C4d-Neo monoclonal antibody, pre-coated on the surface of the microtiter wells. After washing to remove unbound material, a second horseradish peroxidase (HRP) labelled monoclonal antibody, that binds to both allelic variants of C4d (A and B), is added to the well. After incubation for 30 minutes the wells are washed again, and a substrate is added and incubated. The color development is stopped after 30 minutes, and the color is measured in a spectrophotometer. The color is directly proportional to the amount of C4d bound to the well. The amount of C4d is determined by comparison to the color development of the calibrator samples.

 

  • One frame with micro titer wells (12x8) coated with anti-C4d-Neo monoclonal antibody, sealed in a foil pack with a dessiccation bag.
  • 1,5mL Low control (LC). Ready to use
  • 1,5mL High control (HC). Ready to use
  • 2x30 mL Diluent (Dil), red colour. Ready to use.
  • 15 mL Conjugate containing HRP-labelled antibodies to C4d. Ready to use
  • 15 mL Substrate TMB. Ready to use.
  • 15 mL Stop solution (0.5M H2SO4). Ready to use
  • 30 mL Wash solution, 30x concentrated
  • 6 x 1,5mL vials with calibrators containing human recombinant C4d, 0 ng/mL, 5ng/mL, 25 ng/mL, 100ng/mL, 200ng/mL and 400ng/mL. Ready to use

Please note:

All reagents in the kit are ready to use except the wash solution. The reagents should be stored at 2-8°C. Components from different lots shall not be mixed. When stored at 2-8°C the diluted wash solution is stable until the date of expiration of the kit.

Materials or equipment required but not provided

  • Microplate reader with filter 450nm and 620nm
  • Precision pipettes with disposable tips.
  • Washer for strips, absorbent tissue, tubes and a timer.