| Field | Specification |
|---|---|
| Target | |
| Alternative names | Complement C3; C3 and PZP-like alpha-2-macroglobulin domain-containing protein 1; C3; CPAMD1 |
| Gene ID | |
| UniProt # | |
| Reactivity | |
| Applications | |
| Immunogen | Expression system for standard: E.coli; Immunogen sequence: S672-R748 |
| Expression system | |
| Expression region | |
| Assay type | |
| Sample type(s) | Cell Culture Supernatant, Serum, Plasma, Urine |
| Sensitivity | |
| Detection range | |
| Assay time | |
| Storage | |
| Catalog no. (Mfr.) | |
| Main SKU |
Assay Principle
This EZ-Set™ kit supplies a matched capture and detection antibody pair and a standard for building your own sandwich ELISA for human complement C3 in cell culture supernatants, serum, plasma (heparin, EDTA, citrate) and urine. You coat the plate with the capture antibody, add samples and standards, detect with the biotinylated antibody and an avidin–biotin–peroxidase complex, develop with TMB and read at 450 nm. Plates, diluents, wash buffer, TMB and stop solution are not included.
Complement C3 is the central protein of the complement system, where the classical, lectin and alternative pathways converge. Cleavage by C3 convertases produces C3b, which coats pathogens and cells for phagocytosis and forms C5 convertases, and C3a, an anaphylatoxin that promotes inflammation. It is one of the most studied markers of complement activation in immunity and inflammatory disease.
Kit Components
| Mouse anti-human complement C3a monoclonal antibody (Capture Antibody) | 500 µL, 4 µg/mL (recommended dilution 1:100) |
|---|---|
| Biotinylated goat anti-human complement C3a polyclonal antibody (Detection Antibody) | 500 µL (recommended dilution 1:100) |
| Avidin-Biotin-Peroxidase Complex (ABC) | 500 µL (recommended dilution 1:100) |
| Lyophilized recombinant human complement C3a standard | 3 × 10 ng/tube |
Performance Data
| Species | Human |
|---|---|
| Sample types | Cell culture supernatants, serum, plasma (heparin, EDTA, citrate) and urine |
| Assay range | 31.2 pg/mL–2,000 pg/mL |
| Standard | Recombinant human C3 (Ser672–Arg748), expressed in E. coli |
| Cross-reactivity | None detected against related proteins in supplier testing |
Storage & Handling
Store at 4 °C for up to 6 months or −20 °C for up to 12 months. Avoid repeated freeze–thaw cycles. Ships on gel ice and tolerates up to 3 days at room temperature; freeze on arrival.
Safety
Sample data
| Concentration(pg/ml) | 0 | 31.2 | 62.5 | 125 | 250 | 500 | 1,000 | 2,000 |
| O.D. | 0.005 | 0.041 | 0.084 | 0.178 | 0.355 | 0.704 | 1.109 | 2.147 |
Kit components
| Catalog number | Description | Quantity | Storage of opened/reconstituted material |
|---|---|---|---|
| EZ2145-CA | Mouse anti-human complement C3a monoclonal antibody (Capture Antibody) | 500 µl, 4 µg/mL (recommended dilution 1:100) | Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. |
| EZ2145-DA | Biotinylated goat anti-human complement C3a polyclonal antibody (Detection Antibody) | 500 µl (recommended dilution 1:100) | |
| AR1103 | Avidin-Biotin-Peroxidase Complex (ABC) | 500 µl (recommended dilution 1:100) | |
| EZ2145-ST | Lyophilized recombinant human complement C3a standard | 10 ng/tube×3 | Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. |
Materials required but not provided
- Microplate reader in standard size.
- Automated plate washer.
- Incubator.
- Adjustable pipettes and pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection.
- Clean tubes and Eppendorf tubes.
- 96 well microplate (Cat# AR1100)
- Plate Sealers.
- Capture Antibody Diluent: PBS.
- Reagent Diluent: 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered.
- Substrate Solution: Tetramethylbenzidine (Cat# AR1104)
- Stop Solution: 2 N H2SO4 (Cat# AR1105)
- Wash Buffer (PBS and PBS-T). Recipes:
- PBS: 8g NaCl, 0.2g KCl, 1.15g Na2HPO4, 0.2g KH2PO4, adjust the total volume to 1 L with distilled water, pH 7.2-7.4, 0.2 µm filtered.
- PBS-T: 0.1% Tween® 20 in PBS, pH 7.2-7.4.
- *Item 6 – 12 are included in the EZ Set Accessory Kit (EZA001)
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