Calbindin Antibody Staining Protocol for Immunohistochemistry


Description: Calbindin is a calcium-binding protein belonging to the troponin C superfamily. It functions as a buffer for cytoplasmic calcium and is found in the brain, kidney, gut and pancreatic islets. In normal brain, calbindin has been identified in medium sized neurons of the neuropil of the matrix compartment of the striatum, the woolly fibre arrangements of the globus pallidus and the fibre structures of the pars reticula of the substantia nigra. A proportion of dendritic cells within the light zone of germinal centres have also been noted to be positive for calbindin.


Primary Antibody

Name: Calbindin Antibody

Clone: KR6, Mouse anti-Human

Supplier: Novocastra Labs

Catalog Number: NCL-CALBINDIN

Dilution: 1:100 - 1:200 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: Cytoplasmic
Images: Search image

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