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The human complement system is an ancient system for host defense, consisting of > 20 proteins present in plasma and cell surfaces. These proteins interact with each other to opsonize pathogens and induce a series of inflammatory responses to fight infection.
C5 is the fifth component of complement, comprising alpha and beta polypeptide chains linked by a disulfide bond. C5a anaphylatoxin is derived from the alpha chain after cleavage by a convertase upon activation of C5. It is a small polypeptide consisting of 74 amino acids (11 kD). C5a itself is short-lived and cleaved rapidly into a more stable but still biologically active C5a-desArg. Therefore, measurement of C5a-desArg allows reliable quantification of the level of complement activation in the test samples. For convenience, both forms are referred to as C5a in the following manual. Human C5a shares 64% and 62% amino acid sequence identity with mouse and rat, respectively.
C5a is probably the most important complement-derived proinflammatory mediator. It induces the construction of smooth muscle, increases vascular permeability, and causes histamine release. It also stimulates the chemotaxis of polymorphonuclear leukocytes towards sites of inflammation. C5a is believed to play a pivotal role in the pathogenesis of septic shock, adult respiratory distress syndrome, acute pancreatitis and the deleterious effects after myocardial infarction.
The LEGEND MAX™ Human C5a ELISA kit is a Sandwich Enzyme-Linked Immunosorbent Assay (ELISA) with a 96-well strip plate that is pre-coated with a mouse monoclonal anti-human C5a antibody. The Detection Antibody is a biotinylated mouse monoclonal anti-human C5/C5a antibody. This kit is specifically designed for the accurate quantitation of human C5a from serum, plasma, and other biological fluids. This kit is analytically validated with ready-to-use reagents.
- Kit Contents
- Anti-Human C5a Pre-coated 96 well Strip Microplate
- Human C5a Detection Antibody
- Human C5a Standard
- Assay Buffer B
- Wash Buffer (20X)
- Substrate Solution F
- Stop Solution
- Plate Sealers
- Verified Reactivity
- Product Citations
- 2.7 pg/mL
- Standard Range
- 6.25 - 400 pg/mL
- Materials Not Included
- Microplate reader able to measure absorbance at 450 nm
- Adjustable pipettes to measure volumes ranging from 1 µL to 1,000 µL
- Deionized water
- Wash bottle or automated microplate washer
- Log-Log graph paper or software for data analysis
- Tubes to prepare standard dilutions
- Plate Shaker
- Polypropylene vials
- In your LEGEND MAX™ ELISA Kits, there is a step that calls for washing the plates before adding sample. What is the purpose of this step?
We typically use a stabilizer for pre-coated plates. The additional washing step is designed to remove these components before you start the assay. If you do not perform the washing, the effect on assay performance is negligible.
- I have multiple LEGEND MAX™ ELISA kits that I want to run simultaneously. Can I use the same wash buffer for all the kits?
The wash buffer provided in all our LEGEND MAX™ kits is the same and the part numbers on the wash buffer bottles in these kits should be identical. For ELISA MAX™ Deluxe and ELISA MAX™ Standard Sets, we provide a recipe for the wash buffer on each kit’s technical data sheet. This recipe is the same for all ELISA MAX™ sets.
- For some of your ELISA kits, why do my serum samples require dilution with assay buffer?
In some cases, dilution with assay buffer is required to minimize the matrix difference between the samples and the standards to achieve better accuracy.