MRP14 Antibody Staining Protocol for Immunohistochemistry

 

Description: MRP14 (S100A9, Calgranulin B, Heavy Chain of L-1) clone S36.48 identifies the Ca2+ -binding 14kD subunit of the inflammatory L-1 protein complex, also called S100A9 or Calgranulin B. It is useful for the characterization of circulating granulocytes or inflammatory infiltrates of the myelo-monocytic lineage which express MRP14 differently depending on the inflammatory status of the disease.

 

Primary Antibody

Name: MRP14 Antibody

Clone: S36.48, Mouse anti-Human

Supplier: BMA Biomedicals AG

Catalog Number: T-1026

Dilution: 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: Nuclear
Images: Search image

Additional Information:
Tissue Type: CAPAN-1 cell, human lung cancer tissue.
Fixation: Formalin fixed paraffin sections
Positive Control: CAPAN-1 cell, human lung cancer tissue.
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: