 |
|
Cytokeratin Antibody Staining Protocol for Immunohistochemistry
Description: The cytokeratins (CKs) belong to the intermediate filaments, which create a cytoskeleton in almost all eukaryotic cells. In contrast to other intermediate filaments, CKs are made up of a highly complex multigene family of polypeptides with molecular masses ranging from 40 to 68 kDa. CKs are generally held to belong to the most fundamental markers of epithelial differentiation, and until now, 20 distinct CK polypeptides have been revealed in various human epithelia. The CKs can be divided into an acidic type A (class I) and a neutral-basic type B (class II) subfamily.
Primary Antibody
|
Name: Cytokeratin Antibody |
|
Clone: MNF116, Mouse anti-Human |
|
Supplier: Dako |
|
Catalog Number: M0821 |
|
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. |
|
Incubation Time/Temp: 60 min/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 |
| Images: Search image |
Additional Information:
| Species Reactivity: Human |
| Fixation: Formalin fixed paraffin sections, or acetone fixed frozen sections |
| Positive Control: Skin |
| Negative Control: Omit primary antibody, isotype control, 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:
Not need to perform antigen retrieval on acetone fixed frozen sections |
References: