Ubiquitin Antibody Staining Protocol for Immunohistochemistry


Description: This polyclonal antibody detects the presence of intracellular ubiquitinated filamentous inclusions in the periphery of senile plaques and neurofibrillary tangles in Alzheimer's disease and lewy bodies in Parkinsion's disease.


Primary Antibody

Name: Ubiquitin Antibody

Clone: Rabbit polyclonal

Supplier: Dako

Catalog Number: Z-0458

Dilution: 1:500 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: Mouse, rat
Fixation: Formalin fixed paraffin sections
Positive Control: Brain tissue of Alzheimer's or Parkinson's disease
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