Rat Anti-Complement C3
The complement factor C3 consists of an alpha- and a beta-chain. C3 is a central factor in the complement cascade. It is central to the alternative pathway that leads to the C3 convertase C3bBb. The classical and mannose binding lectin activation pathways lead to the C3 convertase C4b2a. These convertases cleave C3 resulting in C3a and C3b. Further degradation leads to formation of the alpha-chain products C3d, C3g and C3c. C3 is an acute phase protein that is produced by a wide range of tissues including renal epithelial cells and hepatocytes.
Applications
Suitable for use in ELISA, Western Blot, Immunoprecipitation, Flow Cytometry, Immunofluorescence and Immunohistochemistry. Other applications not tested.
Recommended Dilutions
ELISA: 0.5ug/well Western Blot: Reduced sample treatment and SDS-PAGE was used. Detects bands at ~110-115kD (alpha chain) and ~70 and ~80KD. Immunofluorescence: Fix in 4% paraformaldehyde in PBS pH 7.4. Vibratome sections of 4uM. Pretreat with 3% hydrogen peroxide for 20 min to quench endogeneous peroxidases. Microwave in antigen unmasking solution for 2-5 min as antigen retrieval. Immunohistochemistry (Frozen): Fix in acetone. Antigen retrieval using 50% formic acid for 5 minutes or microwaved (700W) in antigen unmasking buffer. Optimal dilutions to be determined by the researcher.
Positive Control: MLP-lpr kidney cells
Negative Control: Wild type kidney cells
Storage and Stability
May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 12 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Immunogen
C57BL/6 thymocytes
Form
Supplied as a liquid in PBS, 0.1% BSA, 0.02% sodium azide.
Purity
Purified by Protein G affinity chromatography.
Specificity
Recognizes mouse Complement component C3 and its activation products, C3b, iC3b, C3d and C3dg. Does not crossreact with mouse C4.
