Alpha Internexin Antibody Staining Protocol for Immunohistochemistry

 

Description: Alpha-internexin is a 66kD protein which shares several characteristics with intermediate filament (IF) proteins. In rats, alpha-internexin is often co-purified with intermediate filaments from spinal cord and optic nerve and the protein is axonally transported together with the neurofilament triplet proteins (68, 160 and 200kD) along the length of the optic nerve. Although the distribution of alpha-internexin is similar to the light component neurofilament (68kD), its distribution in rat embryo is far more extensive in the early stages of development.

 

Primary Antibody

Name: alpha-Internexin Antibody

Clone: 2E3, Mouse

Supplier: Novocastra Labs

Catalog Number: NCL-A-INTER

Dilution: 1:50 - 1:100 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: Brain

Fixation: Formalin-fixed paraffin sections

Positive Control: Brain

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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