Ki-67 Antibody Staining Protocol for Immunohistochemistry


Description: Synthetic peptide conjugated to KLH derived from within residues 1200 - 1300 of Human Ki67.


Primary Antibody

Name: Rabbit Anti-Ki67 Antibody

Clone: Rabbit Polyclonal

Supplier: Abcam

Catalog Number: ab15580

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 Chromogen Substrate
Incubation Time/Temperature: 1-3 minutes/room temperature

Reagent: Mayer's Hematoxylin
Staining Time: 30 seconds

Staining Pattern: Nuclear
Images: Search image

Additional Information:
Species Reactivity: Mouse, Rat, Sheep, Rabbit, Horse, Cow, Dog, Human, Pig, Indian Muntjac, Monkey, Chinese Hamster, Marmoset (common), Syrian Hamster
Fixation: Formalin fixed paraffin sections
Positive Control: Skin, tonsil
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