Mitochondria Antibody Staining Protocol for Immunohistochemistry


Description: This antibody recognizes a 60 kDa nonglycosylated protein component of mitochondria found in human cells. It can be used to stain mitochondria in cell, tissue or biochemical preparations and can be used as a marker of the mitochondria in subcellular fractions. Gives mitochondrial staining on all human cell types and does not cross react with rat and mouse tissue. The staining pattern is best described as a spaghetti-like staining pattern in fibroblasts and a perinuclear large speckled pattern in lymphonoid cells. MAB1273 recognizes a 65 kD protein by immunoprecipitation. Antibody produces the stringy, "spaghetti-like" staining patten in the cytoplasm of human cells.


Primary Antibody

Name: Mouse Anti-Mitochondria Antibody

Clone: Mouse monoclonal, clone 113-1

Supplier: Millipore

Catalog Number: MAB1273

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: Cytoplasmic
Images: Search image

Additional Information:
Species Reactivity: Human
Fixation: Formalin fixed paraffin sections
Positive Control: Heart
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