TNF-alpha Antibody Staining Protocol for Immunohistochemistry

 

Description: Specific for natural and recombinant pig TNFα. Does not cross react with recombinant; human TNFα, mouse TNFα, mouse TNFβ, or natural pig TNFβ.

 

Primary Antibody

Name: TNF-alpha Antibody

Clone: Mouse monoclonal

Supplier: Endogen

Catalog Number: MP-390 or MP-391

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/membrane
Images: Search image

Additional Information:
Species Reactivity: Mouse, swine
Fixation: Formalin fixed paraffin sections
Positive Control: Skin, lung
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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