Calretinin Antibody Staining Protocol for Immunohistochemistry


Description: Calbindin D28k and calretinin (also designated CR or 29 kDa Calbindin) are two closely related intracellular calcium-binding proteins belonging to the troponin-C superfamily. Initially isolated from chick retina, calretinin shares 58% identical residues with human calbindin D28k. Calretinin is expressed as a 29 kDa protein in brain and is particularly abundant in auditory neurons with precisely timed discharges. Neurons in the nucleus accumbens containing calretinin all possess nuclear indentations. Calretinin-immunoreactive boutons form asymmetrical and symmetrical synaptic specializations on spines, dendrites and somata. The symmetrical synaptic specializations have medium-sized spiny neurons and contact other Calretinin-immunoreactive somata. Calretinin is widely used as a immunocytochemical marker for mesothelioma.


Primary Antibody

Name: Calretinin Antibody

Clone: Goat anti-Calretinin

Supplier: Santa Cruz Biotechnology

Catalog Number: sc-11644

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: Cytoplasmic
Images: Search image

Additional Information:
Tissue Type: Brain (cerebellum)
Fixation: Formalin fixed paraffin sections
Positive Control: Brain (cerebellum)
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