Home > Protocols > Immunoshistochemical detection of dibromotyrosine Rev.071106
Immunohistochemical detection of dibromotyrosine/dihalogenated tyrpsine/oxidative sress markers/oxidative stress test
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)
oxidative sress markers/oxidative stress test
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.)
[ 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)
oxidative sress markers/oxidative stress test
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