Caspase-2 Antibody Staining Protocol for Immunohistochemistry


Description: Caspases are an extended family of cysteine proteases that play critical roles in apoptosis. Caspases are synthesised as precursors known as pro-caspases and these are converted into mature enzymes by apoptotic signals. Caspase-2 is an early effector in the apoptotic cascade and precedes the activation of caspase-3. It is generally found in the nucleus, but pro-caspase-2 may also be detected in mitochondrial and cytosolic fractions. Lymphoid organs display only weak caspase-2 expression, located in sinusal histiocytes and thymic epithelial cells. Extra-lymphoid caspase-2 reactivity is found in particular organs like the kidney. Bcl-2 oncoprotein may play a central role in inhibiting the caspase-2/caspase-3 cascade.


Primary Antibody

Name: Caspase-2 Antibody

Clone: 10H2, Mouse anti-Human

Supplier: Novocastra Labs

Catalog Number: NCL-CASP-2

Dilution: 1:20 - 1:40 using IHC-TekTM Antibody Diluent (Cat# IW-1000 or IW-1001) to reduce background and unspecific staining and serum blocking step is NOT needed.

Incubation Time/Temp: 60min/room temperature

Antigen Retrieval
Device: IHC-TekTM Epitope Retrieval Steamer Set (Cat# IW-1102)
Buffer/pH value: IHC-TekTM Epitope Retrieval Solution (Cat# IW-1100)
Heat/Cool Temperature: 95-100 ºC/room temperature
Heat/Cool Time: 20 minutes/20 minutes

Detection Methods
Standard Method: ABC Method or LSAB Method
Enhanced Method: Polymeric Methods

Chromogen Substrate
Reagent: DAB
Incubation Time/Temperature: 1-3 minutes/room temperature

Reagent: Mayer's Hematoxylin
Staining Time: 30 seconds

Staining Pattern: Nuclear
Images: Search image

Additional Information:
Tissue Type: Testis
Fixation: Formalin fixed paraffin sections
Positive Control: Testis
Negative Control: Omit primary antibody, isotype control or absorption control
Blocking: 2-5% normal serum to reduce unspecific background staining; 0.5-3% H2O2 to block endogenous peroxidase activity; avidin/biotin to block endogenous biotin activity if necessary