Beta Galactosidase Antibody Staining Protocol for Immunohistochemistry


Description: Beta galactosidase is a a metalloenzyme that that splits lactose into glucose and galactose and is coded by a gene (lac z) in the lac operon of Escherichia coli.


Primary Antibody

Name: Beta galactosidase Antibody

Clone: Rabbit polyclonal

Supplier: Abcam

Catalog Number: ab616

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: 60min/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: Nulcear
Images: Search image

Additional Information:
Tissue Type: Tissue or cells expressing LacZ or Transgenic mice expressing nuclear beta-galactosidase
Fixation: Formalin-fixed paraffin sections, or paraformaldehyde fixed frozen sections
Positive Control: Tissue or cells expressing LacZ or Transgenic mice expressing nuclear beta-galactosidase
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


There is no need to perform antigen retrieval on acetone fixed frozen sections



The dilution of this antibody varies depending on tissue type, fixation, pretreatment, etc