Cytokeratin 7 Antibody Staining Protocol for Immunohistochemistry

 

Description: The antibody reacts with cytokeratin 7 which has been found in columnar and glandular epithelium of the lung, cervix, breast, bile ducts and larger collecting ducts of the kidney. Occasionally staining of blood vessel cell walls, particularly endothelial cells, may be observed.

 

Primary Antibody

Name: Cytokeratin 7 FITC conjugate Antibody

Clone: LP5K, Mouse anti-Human

Supplier: Chemicon

Catalog Number: CBL194F

Dilution: 1:50 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 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) or IHC-TekTM Proteinase K Solution (Cat# IW-1101)
Heat/Cool Temperature: 95 ºC - 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:
Tissue Type: Skin, lung, breast, kidney.
Fixation: Formalin fixed paraffin sections, or acetone fixed frozen sections
Positive Control: Skin, lung, breast, kidney.
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:

No need to perform antigen retrieval on frozen sections

 

References: