MIP-3a Antibody Staining Protocol for Immunohistochemistry

 

Description: This antibody was produced from a hybridoma resulting from the fusion of a mouse myeloma with B cells obtained from a rat immunized with purified, E. coli-derived, recombinant mouse macrophage inflammatory protein 3 alpha (rmMIP-3a). MIP-3a is a member of the CC chemokine superfamily and has been designated CCL20.

 

Primary Antibody

Name: MIP-3a Antibody

Clone: 114908, Rat anti-Mouse

Supplier: R&D Systems

Catalog Number: MAB7601

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 staining of macrophage
Images: Search image

Additional Information:
Tissue Type: Small intestine
Fixation: Formalin fixed paraffin sections
Positive Control: Small intestine
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: