Activation of procaspase-9 by Apaf-1 in the cytochrome c/dATP- dependent pathway requires proteolytic cleavage to generate the mature caspase molecule. Deletion of the Apaf-1 WD-40 repeats makes Apaf-1 constitutively active and capable of processing procaspase-9 independent of cytochrome C and dATP. Apaf-1-mediated processing of procaspase-9 occurs at Asp-315 by an intrinsic autocatalytic activity of procaspase-9 itself. Apaf-1 can form oligomers and may facilitate procaspase-9 autoactivation by oligomerizing its precursor molecules. Once activated, caspase-9 can initiate a caspase cascade involving the downstream executioner caspases- 3, -6, and -7.
Applications:
Suitable for use in Western Blot and Immunohistochemistry. Other applications not tested.
Recommended Dilution:
Western Blot: 2.5-5ug/ml
Immunohistochemistry: 1:100 for 10min at RT
Staining of formalin-fixed tissues requires boiling tissue sections in 10mM citrate buffer, pH 6.0, for 10min followed by cooling at RT for 20min
Optimal dilutions to be determined by the researcher.
Storage and Stability:
May 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 at least 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
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