Eotaxin (EOT) Antibody Staining Protocol for Immunohistochemistry

 

Description: This antibody has been selected for its ability to neutralize the biological activity of rmEotaxin and for use as a capture antibody in sandwich ELISAs. It will also neutralize the biological activity of rhEotaxin at a 2-3 fold higher IgG concentration.

 

Primary Antibody

Name: Eotaxin Antibody

Clone: Goat anti-Mouse

Supplier: R&D Systems

Catalog Number: AF-420-NA

Dilution: 1:20 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 minutes/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 ºC - 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: Heart
Fixation: Formalin fixed paraffin sections, or acetone fixed frozen sections
Positive Control: Heart
Negative Control: Omit primary antibody, isotype control or 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

Notes:

No need to perform antigen retrieval on frozen sections

 

References: