MDR Antibody Staining Protocol for Immunohistochemistry


Description: Cells selected for resistance to a single cytotoxic drug may become cross-resistant to a broad range of drugs with different structures and cellular targets. This phenomenon is called multiple drug resistance. The multiple drug resistance (Mdr) gene encodes a 170 kDa glycoprotein (p-glycoprotein) that is a member of a highly conserved superfamily of ATP-binding cassette transport proteins. p170 glycoprotein functions as an energy-dependent efflux pump for structurally diverse agents ranging from ions to peptides. p170 glycoprotein has been implicated in the development of the multiple drug resistance observed in human cancer cells following prolonged chemotherapy. The classic form of Mdr is associated with an increase in p glycoprotein, but not all cases of Mdr can be attributed to a rise in the levels of p-glycoprotein. In cell lines not expressing increased levels of p-glycoprotein, researchers found an increase in the level of a novel protein designated as Mdrassociated protein (MRP).


Primary Antibody

Name: MDR (Multiple Drug Resistance, H-241) Antibody

Clone: Rabbit polyclonal

Supplier: Santa Cruz Biotechnology

Catalog Number: sc-8313

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

Reagent: Mayer's Hematoxylin
Staining Time: 30 seconds

Staining Pattern: Membrane
Images: Search image

Additional Information:
Tissue Type: Colon cancer
Fixation: Formalin fixed paraffin sections
Positive Control: Colon cancer
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