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Cytokeration 5&8 Antibody Staining Protocol for Immunohistochemistry
Description:
Cytokeratins are a subfamily of
intermediate filament proteins and are characterized by a remarkable
biochemical diversity, represented in epithelial tissues by at least
20 different polypeptides. They range in molecular weight from
between 40 kDa and 68 kDa and isoelectric pH between 4.9 - 7.8, The
individual cytokeratin polypeptides are designated 1-20. The various
epithelia in the human body usually express cytokeratins which are
not only characteristic of the type of epithelium, but also related
to the degree of maturation or differentiation within an epithelium.
Cytokeratin subtype expression patterns are used to an increasing
extent in the distinction of different types of epithelial
malignancies.
Primary Antibody
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Name: Cytokeratin 5&8 Antibody |
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Clone: Mouse monoclonal |
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Supplier: Chemicon |
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Catalog Number: MAB3228 |
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Dilution: 1:200 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: 60 minutes/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/membrane |
| Images: Search image |
Additional Information:
| Tissue Type: Skin |
| Fixation: Formalin fixed paraffin sections, or acetone fixed frozen sections. |
| Positive Control: Skin, lung, pancreas |
| 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 frozen sections. |
References: