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Protocols > Immunoshistochemical detection of dibromotyrosine |
Rev.071106 |
| This is an application example for anti dibromotyrosine / di-halogenated tyrosine monoclonal antibody.
Experimental conditions should be settled by each user. |
| Fixation: |
Fix by neutral formalin solution. |
| Deparaffinization: |
Deparaffinization (In case of paraffin embedded specimens).Deparaffin with xylene and ethanol.
(Optional: antigen retrieval.) |
| Blocking: |
Normal rabbit serum (DAKO) in 1%BSA/PBS for 30 minutes. |
| Primary antibody: |
10 µg/mL of Anti dibromotyrosine antibody (clone 3A5) at 4 °C, overnight. |
| Detection: |
Biotinated anti mouse IgG (DAKO)
Avidin-Biotin-Complex (ABC) reagent: ABC-AP Kit (Vector)
Chromogen: Alkaline Phosphatase Kit (Vector) |
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Antigen retrieval: Please try if staining is too weak or if background is too high.
Microwave treatment for 5 minutes in 0.01M citrate buffer (pH6.0).
(Technical information: kindly provided by Dr. Yoji Kato, University of Hyogo.) |
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| [ Reference ] |
| 1) |
Kato Y, Kawai Y, Morinaga H, Kondo H, Dozaki N, Kitamoto N, Osawa T.
Immunogenicity of a brominated protein and successive establishment of a monoclonal antibody to dihalogenated tyrosine.
Free Radic Biol Med. 38(1), p24-31(2005) |
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