Keratin 15 Antibody Staining Protocol for Immunohistochemistry

 

Description: Keratin 15 is a type I keratin without a defined type II partner. Keratin 15 is expressed primarily in the basal keratinocytes of stratified tissues, including the fetal epidermis and fetal nail. Expression of keratin 15 is downregulated in some hyperproliferating situations, such as psoriasis and hypertrophic scars. Because keratinocytes in psoriasis and hypertrophic scars are activated, it is suggested that keratin 15 expression is not compatible with keratinocytes activation and the keratin 15 gene is downregulated to maintain the activated phenotype.

 

Primary Antibody

Name: Keratin 15 Antibody

Clone: LHK15, Mouse monoclonal

Supplier: Lab Vision

Catalog Number: MS-1068

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:
Tissue Type: Skin
Fixation: Formalin fixed paraffin sections. Not tested on 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

 

References: