0.2 µm filtered in phosphate-buffered solution, pH 7.2, containing no preservative. Endotoxin level is <0.1 EU/µg of the protein (<0.01 ng/µg of the protein) as determined by the LAL test.
Preparation
The LEAF™ (Low Endotoxin, Azide-Free) antibody was purified by affinity chromatography.
Concentration
1mg/ml
Recommended Usage
Each lot of this antibody is quality control tested by ELISA assay. For ELISA capture applications, a concentration range of 2-6 µg/ml is recommended. To obtain a linear standard curve, serial dilutions of C3a recombinant protein ranging from 39 to 2500 pg/ml are recommended for each ELISA plate. It is recommended that the reagent be titrated for optimal performance for each application.
Applications
ELISA Capture - Quality tested IP, WB, Neut - Reported in the literature
Application Notes
The K13/16 antibody reacts with human C3a, C3a(desArg) and C3. The K13/16 antibody can neutralize the bioactivity of natural or recombinant C3a.
ELISA Capture: The purified K13/16 antibody is useful as the capture antibody in a sandwich ELISA assay, when used in conjunction with the biotinylated D17/1 antibody (Cat. No. 518002) as the detecting antibody.
Neutralization: The LEAF™ Purified antibody (Endotoxin <0.1 EU/μg, Azide-Free, 0.2 μm filtered) is recommended for neutralization of human C3a/C3a(desArg)/C3 bioactivity.
Application References
1. Oppermann M, et al. 1991. Kidney Int. 40:939947. 2. Oppermann M, et al. 1992. Immunopharm. 24:119134. 3. Elsner J, et al. 1994. Blood 83:3324.
Description
C3, complement component 3, is a 195 kD glycoprotein heterodimer that is linked by disulfide bonds. It is the fourth complement component to react in the classical pathway and it is also a reactant in the alternative complement pathway. C3 convertase splits C3 into C3b and C3a. C3b can bind covalently, via its reactive thioester, to cell surface carbohydrates or immune aggregates. Macrophages and neutrophils have receptors for C3b. C3a is released into the surrounding fluids. It can bind to receptors on basophils and mast cells triggering them to release their vasoactive contents (e.g., histamine). Because of the role of these materials in anaphylaxis, C3a is called an anaphylatoxin. In blood, the C3a fragment is rapidly desarginated by serum carboxypeptidase N enzyme to generate C3a desArg. Both C3a and C3a desArg are not glycosylated.
Storage
The antibody solution should be stored undiluted at 4°C. This LEAF™ solution contains no preservative; handle under aseptic conditions.