Description
The I52-278 monoclonal antibody reacts with the mouse C5a protein. Clone I52-278 hybridoma was generated by the fusion of spleen cells from rats immunized with recombinant mouse C5a protein. Anaphylatoxin C5a is a bioactive cleavage product released from plasma component C5 during complement activation and is involved in mediation of a variety of cellular immune responses, as well as being potent pro-inflammatory agents. The release of this cleavage product is a reliable indicator of in vivo or in vitro complement activation.
The biotinylated I52-278 antibody is useful as a detector antibody for a sandwich ELISA that measures mouse C5a protein levels. Purified I52-1486 as capture antibody (Cat. No. 558027) can be paired with the biotinylated I52-278 anti-mouse C5a as the detection antibody, using purified recombinant mouse C5a protein (Cat. No. 622597) as standard. The biotinylated I52-278 antibody should be titrated between 0.25-2 µg/ml to determine its optimal ELISA detection concentration. To obtain linear standard curves, doubling dilutions of purified mouse C5a protein ranging from 156 pg to 10 ng/ml are recommended for inclusion in each ELISA plate. PBS with 10%FCS is recommended as the blocking buffer. Addition of FUT-175 (Futhan), Cat. No. 552035, to plasma samples at the time of sample collection provides additional protection from ex-vivo activation, and therefore ensures more accurate measurements that reflect the circulating levels of complement activation products. For specific methodology and ELISA buffer recipes, see Immune Function Handbook which is posted on the website at www.bdbiosciences.com. This ELISA antibody pair shows no cross-reactivity with the following: recombinant mouse C3a, native human, and rat C5a.
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with biotin under optimum conditions, and unreacted biotin was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
3. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
