Immunohistochemical analysis of paraffin-embedded E14 mouse embryo using Cleaved Caspase-3 (Asp175) (D3E9) Rabbit mAb #9579 (left) or Rabbit (DA1E) mAb IgG XP® SignalStain® Isotype Control #12960 (right).
Immunohistochemical analysis of paraffin-embedded Kelly xenograft using Cleaved Caspase-3 (Asp175) (D3E9) Rabbit mAb #9579 (left) or Rabbit (DA1E) mAb IgG XP® SignalStain® Isotype Control #12960 (right).
|Product Includes||Quantity||Storage Temp|
|Cleaved Caspase-3 (Asp175) (D3E9) Rabbit mAb 9579||100 µl||-20°C|
|Rabbit (DA1E) mAb IgG XP® SignalStain® Isotype Control||100 µl||-20°C|
|Normal Goat Serum 5425||2.4 ml||-20°C|
|SignalStain® Boost IHC Detection Reagent (HRP, Rabbit) 8114||12 ml||4°C|
|SignalStain® DAB Diluent||12 ml||4°C|
|SignalStain® DAB Chromogen Concentrate||360 µl||4°C|
|Tris Buffered Saline with Tween® 20 (TBST-10X) 9997||2.4 ml||RT|
|SignalStain® Antibody Diluent 8112||25 ml||4°C|
NOTE: Additional 10X TBST will be required for washes.
NOTE: Do not allow slides to dry at any time during this procedure.
posted July 2013
revised June 2016
Protocol Id: 64
SignalStain® Apoptosis (Cleaved Caspase-3) IHC Detection Kit allows the detection of activated caspase-3 in formalin-fixed paraffin-embedded human and mouse tissue samples. Cleaved Caspase-3 (Asp175) (D3E9) Rabbit mAb is detected by the polymer based, HRP-conjugated SignalStain® Boost IHC Detection Reagent in combination with SignalStain® DAB Diluent and Chromogen Concentrate. Also included is a concentration-matched rabbit monoclonal IgG control to verify the specificity of staining.
This combination of reagents provides a sensitive and specific means of detecting apoptotic events in tissue samples.
SignalStain® Apoptosis (Cleaved Caspase-3) IHC Detection Kit detects endogenous levels of the activated caspase-3 large fragment (17/19 kDa) resulting from cleavage adjacent to Asp175. This antibody does not recognize full-length caspase-3 or other cleaved caspases. This kit was developed for and is recommended for immunohistochemistry only.
Cleaved Caspase-3 (Asp175) (D3E9) Rabbit mAb is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Asp175 of human caspase-3 protein. Rabbit (DA1E) mAb IgG XP® SignalStain® Isotype Control is not directed against any known antigen. It functions as an isotype control for rabbit IgG monoclonal antibodies.
Caspase-3 (CPP-32, Apoptain, Yama, SCA-1) is a critical executioner of apoptosis, as it is either partially or totally responsible for the proteolytic cleavage of many key proteins, such as the nuclear enzyme poly (ADP-ribose) polymerase (PARP) (1). Activation of caspase-3 requires proteolytic processing of its inactive zymogen into activated p17 and p12 fragments. Cleavage of caspase-3 requires the aspartic acid residue at the P1 position (2).
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