More info
Assay Range | 15.6--1000 pg/mL |
Sensitivity | 10.0 pg/mL |
Size | 96T |
Storage | Store at 2 - 8ºC. Keep reconstituted standard and detection Ab at -20 ºC |
Assay Principle | Sandwich ELISA |
Sample volume | 100 µL final volume, dilution factor varies on samples. |
Detection Method | Chromogenic |
Kit Components
1. Recombinant Mouse C3a standard | 2 vials. |
2. One 96-well plate precoated with anti-Mouse C3a Ab | 1. |
3. Sample diluent buffer | 12 ml- 2. |
4. Detection antibody | 130 µL, dilution 1:100. |
5. Streptavidin-HRP | 130 µL, dilution 1:100. |
6. Antibody diluent buffer | 12 ml x1. |
7. Streptavidin-HRP diluent buffer | 12 ml x1. |
8. TMB developing agent | 10 ml x1. |
9. Stop solution | 10 ml x1. |
10. Washing solution (20x) | 25 ml x1. |
Background
Complement C3 is a naturally glycosylated (approximately 2.7%) polypeptide containing two disulfide-linked chains. C3 plays a key role in the activation of all three pathways of complement activation. Initiation of each pathway generates proteolytic enzyme complexes (C3 convertases) which are bound to the target surface. These enzymes cleave a peptide bond in C3 releasing the anaphylatoxin C3a and activating C3b. C3a binds the G-protein coupled C3a receptor (C3aR) on myeloid peripheral blood leukocytes,and on activated lymphocytes, endothelial and internal organ epithelial cells. It activates mast cells and neutrophils, triggering robust mast cell degranulation in airways during asthmatic allergen challenges. It enhances LPS induced prostaglandin, cytokine and chemokine secretion by macrophages and other cells. It assists in Th2-type inflammatory reactions and stimulates smooth muscle contraction and leukocyte chemotaxis. Many of the biological functions of C3a are similar to those of C5a, but C3a is approximately 10- to 20-fold less active per microgram than C5a. C3b is linked to the target through a covalent ester bond. Surface-bound C3b is necessary in all three pathways for efficient activation of C5 and formation of C5b-9 complexes that lyse the target cell membrane.