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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
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Name: Cellular Apoptosis Susceptibility Protein (CAS) Antibody |
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Clone: 30F12, Mouse anti-Human |
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Supplier: Novocastra Labs |
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Catalog Number: NCL-CAS |
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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. |
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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:
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There is no need to perform antigen retrieval for acetone fixed frozen sections |
References: