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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
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Name: Calbindin Antibody |
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Clone: KR6, Mouse anti-Human |
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Supplier: Novocastra Labs |
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Catalog Number: NCL-CALBINDIN |
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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. |
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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 |
Counterstain
| Reagent: 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: