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Beta-Tubulin III Antibody Staining Protocol for Immunohistochemistry
Description: Beta-Tubulin III (Clone Tuj1). This antibody was raised against microtubules derived from rat brain and recognizes the peptide epitope, CEAQGPK. It is well characterized and highly reactive to neuron specific Class III β-tubulin (βIII). Tuj1 does not identify β-tubulin found in glial cells.
Primary Antibody
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Name: Beta Tubulin III Antibody |
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Clone: Tuj1, Mouse monoclonal |
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Supplier: Covance |
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Catalog Number: MMS-435P |
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Dilution: 1:1200 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: 60 min/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: Neuroal body and axons |
| Images: Search image |
Additional Information:
| Species Reactivity: Mouse, rat |
| Fixation: Formalin fixed paraffin sections |
| Positive Control: Brain (cerebral cortex) |
| 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: