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 人GTP酶活化蛋白(GAP)ELISA 试剂盒
  • 人GTP酶活化蛋白(GAP)ELISA 试剂盒
  • 人GTP酶活化蛋白(GAP)ELISA 试剂盒

    参考价格: ¥3000.00
    品  牌:USA
    产品型号:RC01020B (人GTP酶活化蛋白(GAP)ELISA Kit )
    所在地区:上海
    上架时间:2012年08月01日
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    详细说明

    RC01020B 人GTP酶活化蛋白(GAP)ELISA 试剂盒  人GTP酶活化蛋白(GAP)ELISA Kit
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    ASSAY METHOD
     Before use, mix all reagents thoroughly without making foam.
     Determine the number of microwell strips required to test the desired number of samples,plus appropriate number of wells needed for running blanks standards. Each sample, standard and blank should be assayed in duplicate. Remove sufficient microwell strips from the pouch.
     Add 50ul of standard diluent to standard wells B1,B2,  C1,C2,  D1,D2,  E1, E2,  F1,F2. Reconstitute standard vial with the appropriate volume as described in the chapter reagents preparation. Preparation. Pipet 100ul of standard into wells A1 and A2 (see plate scheme below). Transfer 50ul from A1 and A2 to B1 and B2 wells. Mix the contents by repeated aspirations and ejections. Take care not to scratch the inner surface of microwells. Repat this procedure from the wells B1,B2 to wells C1,C2 and from wells C1,C2 to D1,D2 and so on creating two parallel rows of p16 standard dilutions ranging,Add 50ul of standard diluent to the bland wells.
     Dilute samples 1:1 distribing 50ul of sample into 50ul of dilluent ,Add 50ul of diluted sample to wells..
     Add 50ul of diluted biotinylated anti-p16 to all wells.
     Cover with a plate vover and incubate for 1 hour at 37℃.
     Remove the cover and wash the plate as follows: ⑴ aspirate the liquid from each well, ⑵ dispensse 0.3ml of washing solution into each well. ⑶ Aspirate again the contet of each well after 0.5 minute. ⑷ Repeat steps ⑵ and ⑶ three times.
     Distribute 80ul of streptavidin-HRP solution to all wells, including blank wells.
     Cover and incubate 30 min at 37℃.
     Remove the cover and empty wells, Wash microwell strips according to step, Proceed immediately to the next step.
     Add 50ul Substrate A and Substrate B to each well。Incubate for 10 min at 37℃。
     The enzyme-substrate reaction is stopped by quickly pipetting 50ul of H2SO4. stop reagent into each well, including the blank wells, to completely and uniformly inactivate the enzyme. Results must be red immediately after the addition of H2SO4.
     Read absorbance of each well on a spectrophotometer using 450nm as the primary wavelength and optionally 620nm (610nm to 650nm is acceptable ) as the reference wavelength.

     

    SUGGESTED PLATE SCHEME
     Standard
    concentrations(ng/ml) 
    A 80 80 sample sample sample sample sample sample sample sample sample sample
    B 40 40 sample sample sample sample sample sample sample sample sample sample
    C 20 20 sample sample sample sample sample sample sample sample sample sample
    D 10 10 sample sample sample sample sample sample sample sample sample sample
    E 5.0 5.0 sample sample sample sample sample sample sample sample sample sample
    F 2.5 2.5 sample sample sample sample sample sample sample sample sample sample
    G 0 0 sample sample sample sample sample sample sample sample sample sample
    H sample sample sample sample sample sample sample sample sample sample sample sample

    LIMITATIONS OF THE PROCEDURE
    Do not extrapolate the standard curve beyond the max  standard curve point. The dose-response is non-linear in this region and good accruacy is difficult to obtain.

    CALCULATION OF RESULTS
    The minimum detectable concentration in this assay is estimated to be 0.1ng/ml