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.\n\nApplications:\nSuitable for use in ELISA, Western Blot, Immunoprecipitation, Flow Cytometry, Immunofluorescence and Immunohistochemistry. Other applications not tested. \n\nRecommended Dilution:\nELISA: 0.5ug/well\nWestern Blot: Reduced sample treatment and SDS-PAGE was used. Detects bands at ~110-115kD (alpha chain) and ~70 and ~80KD.\nImmunofluorescence: 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. \nImmunohistochemistry (Frozen sections): fix in acetone. Antigen retrieval using 50% formic acid for 5 min or microwaved (700W) in antigen unmasking buffer.\nImmunohistochemistry (paraffin): Pretreat free-floating brain tissue with 50% formic acid for 5 min. or microwave (700W) in antigen unmasking solution for 2-5min. Block endogenous persoxidase by treatment with 3% H2O2 in PBS 20 min at 25 degrees C. Block 2hrs, 2% BSA in PBS.\nOptimal dilutions to be determined by the researcher.\n\nPositive Control: MLP-lpr kidney cells\n\nNegative Control: Wild type kidney cells\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20 degrees C. Aliquots are stable for 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

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