Keratin 7 Antibody Staining Protocol for Immunohistochemistry

 

Description: Cytokeratin 7 is a basic cytokeratin which is found in most glandular and transitional epithelia but not in the stratified squamous epithelia. Keratin 7 is expressed in the epithelial cells of ovary, lung, and breast but not of colon, prostate, or gastrointestinal tract. Antibody to cytokeratin is useful in distinguishing ovarian carcinomas (keratin 7+) from colon carcinomas (keratin 7).

 

Primary Antibody

Name: Keratin 7 Antibody

Clone: K72.7, Mouse anti-Human

Supplier: Lab Vision

Catalog Number: MS-283-P

Dilution: 1:1000 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:
Tissue Type: Skin
Fixation: Formalin fixed paraffin sections. Not tested on frozen sections.
Positive Control: Lung, ovary, cervix, endometrium, and breast carcinoma, 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

 

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