Cellular Apoptosis Susceptibility Protein (CAS) Antibody Staining Protocol for Immunohistochemistry

 

Description: Cellular apoptosis susceptibility protein, the product of the CAS gene, is associated with microtubules and the mitotic spindle. CAS is the human homologue of the yeast chromosome-segregation gene, CSE-1. The molecular mechanism or function by which CAS is associated with cell proliferation and apoptosis is not yet fully understood. High expression of cytoplasmic CAS protein appears to correlate with proliferation of lymphoid cells. CAS expression may be useful in the identification of proliferating cells and in the elucidation of the function of cellular apoptosis.

 

Primary Antibody

Name: Cellular Apoptosis Susceptibility Protein (CAS) Antibody

Clone: 30F12, Mouse anti-Human

Supplier: Novocastra Labs

Catalog Number: NCL-CAS

Dilution: 1:40 - 1:80 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

Counterstain
Reagent: Mayer's Hematoxylin
Staining Time: 30 seconds

Results:
Staining Pattern: Cytoplasmic/nuclear
Images: Search image

Additional Information:
Tissue Type: Tonsil
Fixation: Formalin fixed paraffin sections, or acetone fixed frozen sections
Positive Control: Tonsil
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

Notes:

There is no need to perform antigen retrieval for acetone fixed frozen sections

 

References: