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                              千赢国际真人

     

    SPECIFICATION AND INSTRUCTION

    [Product Name]

    千赢国际真人

    [Package]

    20 samples per kit

    [Intended Use]

    For detecting tumor protein p185erbB-2/HER-2 provide the basis of diagnosis in Breast cancer and ovarian cancer .

    [Test Principle]

    erbB-2/HER-2 is a oncogene, the protein of 185kD(p185),the positive expression of p185  becomed  a significant tumor marker of international generally accepted. This kit uses self-developed Anti-p185erbB-2/HER-2 Monoclonal antibody, and perform Biotin-Avidin IHC , cancer membrane Stained yellow means positive .

    [Units of Kit]

    (1)  Solution A        5.0ml      3% H2O2

    (2)  Solution B        1.2ml      Blocking Solution

    (3)  Solution C        1.2ml      Anti-p185erbB-2/HER-2Monoclonal antibody

    (4)  Solution D        1.2ml      Biotinylated secondary  antibody                            

    (5)  Solution E        1.2ml      Avidin-labeled HRP                        

    (6)  DAB             prepared before use

    (7)  positive tissue slice  (Purchased youself) : tissue slice of p185erbB-2/HER-2 positive                              

    (8)  Instruction.

    [Storage]

    Stored at 2 ~ 8℃ for 10 months

    [Equipment]

    Common light microscope

    [Sample]

    Tissue slice

    [Test method]

    (1) Paraffin Sections: Trim paraffin blocks as necessary and cut at 4 um (5 um is commonly used).And the frozen sections,should have been used fixation for frozen tissue sections is to immerse the slides in pre-cooled acetone 4℃ for 10 min;

    (2) Dry sections, melting Paraffin:Dry sections in incubator for at least 15 min. at -90℃;

    (3)Deparaffinization/Rehydration:Hydrate with Xylene thanol and rinse in distilled water,then can perform Immunohistochemistry;

    (4) Rinse in PBSH7.4,0.01M Phosphate buffer or 3x5min;

    (5) Allow the slide to drain, Shake off excess Liquid with a brisk motion and carefully wipe each slide around the Sections, Dropping A solution cover the tissue Sections ,incubate

    10 min at Room temperature. Shake off excess Liquid, Rinse in PBS for 3x5min;

    (6) Allow the slide to drain, Shake off excess Liquid with a brisk motion and carefully wipe each slide as before ,apply B solution cover the tissue Sections ,incubate 10 min at Room temperature;

    (7) Allow the slide to drain, Shake off excess Liquid with a brisk motion and carefully wipe each slide as before, apply C solution cover the tissue Sections, apply B solution cover the tissue Section. ,incubate for 1 hour at 37℃ or overnight at 4℃chamber. Rinse in PBS for 3x5min; Allow the slide to drain, Shake off excess Liquid with a brisk motion and carefully wipe each slide as before, apply B solution cover the tissue Section, incubate 10 min at Room temperature, Rinse in PBS for 3x5min;

    (8)  Allow the slide to drain, Shake off excess Liquid with a brisk motion and carefully wipe each slide as before ,apply E solution cover the tissue Sections ,incubate 10 min at Room temperature, Rinse in PBS for 3x5min;.

    (9)  Prepared DAB solution when it been used, dissolve 5mgDAB in 10ml of PBS,and apply A solution 100µl, (uint desired color reaction is observed when monitored with  the  Microscope);

    (10) Rinse in distilled water ,or can do karyotin again when necessary; Dehydrate through Gradient ethanol, Clear in xylene, Coverslip with Gum.

    [Reference Result]

    Reference value: compared to negative tissue slice ,the positive tissue slice should have 5% cancer membrane Stained yellow, immune reactions were continuous and linear around cell.

    [Interpretation of the Results Identified]

    Negative: the tissue Sections without stained, <5% cancer membrane Stained yellow, Staining intensity is Low.  

    Low positive (±-+) :> 5% cancer membrane Stained yellow, immune reactions were not continuous and linear around cell . Staining intensity is Low,part of cell Stained.

    Moderate positive(++): > 5% cancer membrane Stained yellow, immune reactions were continuous and linear around cell        

    High positive(+++): > 5% cancer membrane Stained dense yellow, immune reactions were continuous and linear around cell.++-+++ means over-expression

    Precaution: The stained Cytoplasm can not be judged for Specific staining .

    [Limitations of Test Methods]

    Only for tissue slice

    [Product Performance]

    Comparison with similar products, Correlation coefficient0.654<0.0001

    [Precaution]

    (1) By use this kit,the  Sections without prior enzyme digestion or antigen rerival.

    (2) All solutions applied should  cover the tissue Sections

    (3) Room temperature mean 20 �, Bath at 37 � better  

    (4) pH7.4, 0.01MPBS and DAB need Prepared before use by youself

    [Manufactured by]

    Anhui Anke Biotechnology (Group) Co., Ltd.

    [Address]

    Ankebio Buildings, 669, Changjiang Road, West ,Hefei, 230088, P.R. China.

    Tel: +86-551-65319890, 65318811

    Fax:+86-551-65319895

    Website: www.ankebio.com

    E-mail: info@ankebio.com

    [Medical Device Manufacturing Enterprise License Number] WSYJX 20070120

    [The registration Number of Medical Devices] GSYJX(Z)Z2009N3400543

    [Standard Number]  YBS00392004

    [Instruction Approval and Modified day] July 21st ,2009

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