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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
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Name: Keratin 15 Antibody |
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Clone: LHK15, Mouse monoclonal |
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Supplier: Lab Vision |
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Catalog Number: MS-1068 |
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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 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: