Methods & Techniques
For Histologists and Immunohistochemists
 

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

Incubation Time/Temp: 60min/room temperature


Antigen Retrieval
Device: Steamer
Buffer/pH value: Citrate buffer/pH 6.0
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

Counterstain
Reagent: Gill's Hematoxylin or Mayer's Hematoxylin
Staining Time: 30 seconds

Results:
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

 

References: