Assay kit contains reagents optimized for immunocytological detection of cells undergoing DNA replication. The thymidine analog, 5-Bromo-2-deoxyuridine (BrdU), is added to the cell growth medium just prior to fixing and staining the cells with an antibody to BrdU, which detects the thymidine analog in DNA. DNA denaturation is not required, enabling co-staining for other proteins of interest. A bridging antibody (for signal amplification), a fluorophore-conjugated tertiary antibody and wash buffer are included for convenience.
Quantity:
Minimum of 40 DNA replication assays.
Kit Components:
D3949-50A: BrdU antibody 4x2.5ml.
D3949-50B: IgG (CyTM2) 1x100ug
D3949-50C: IgG 1x100ug
D3949-50D: BrdU (5-Bromo-2'-deoxyuridine) 1x25mg
D3949-50E: TBS (20X) 1x50ml
Quality Control:
Immunocytochemistry:
HeLa cells incubated for 30 minutes with 10uM BrdU and fixed for 30 minutes with 95% ethanol, 5% glacial acetic acid showed positive immunostaining for BrdU using a 1:4 dilution anti-BrdU, clone BU-1, 5ug/ml Goat anti-Mouse bridging antibody and a 1:100 dilution of anti-goat CyTM2 conjugate.
Storage and Stability:
Store D3949-50A as a powder at -20 degrees C, as a liquid at 4 degrees C. Store D3949-50B, D3949-50C and D3949-50D at -20 degrees C, in the dark. Store D3949-50E at 4 degrees C. Stable for 6 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
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