人磷脂酰肌醇蛋白聚糖3(GPC-3)检测试剂盒齐一生物科技(上海)有限公司销售电话:021-60348496.www.qiyibio.com:欢迎来电询问原料价格及销售库存.绝不生产销售劣质产品,欢迎广大消费者监督是我公司重点推广产品,我公司有专业的人员进行全程指导,请放心购买,发货时均会附上质检报告单.使用说明书和推荐用法用量,提供正规发票.齐一生物科技(上海)有限公司客服热线:021-60348467 60348496.Web:www.qiyibio.com
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FOR RESEARCH USE ONLY
Drug Names
Generic Name:人磷脂酰肌醇蛋白聚糖3(GPC-3)检测试剂盒 This kit can be used for determination of serum, plasma and liquid samples Organization Content.
The experimental principle:
The product levels were measured in samples of the kit by double antibody sandwichmethod. The product with the purified antibody coated microtiter plate, made of solid phase antibody, to package is the product antigen monoclonal antibodies are then added to the micropores, the product and then with HRP labeled antibody binding, the formation of antibody - antigen - antibody complex enzyme label, after thorough washing with TMB chromogenic substrate. TMB in the HRP enzyme catalytic conversion into the blue, and in the action of acid into the final yellow. This product is positively related to the depth of color and in the samples. Instrument measured absorbance in the 450nm wavelength with ELISA (OD), the product concentration in the samples was calculated by standard curve.
Materials provided with the kit
Materials provided with the kit 48determinations 96 determinations Storage
User manual 1 1
Closure plate membrane 2 2
Sealed bags 1 1
Microelisa stripplate 1 1 2-8℃
Standard:360ng/L 0.5ml×1 bottle 0.5ml×1 bottle 2-8℃
Standard diluent 1.5ml×1 bottle 1.5ml×1 bottle 2-8℃
HRP-Conjugate reagent 3ml×1 bottle 6ml×1 bottle 2-8℃
Sample diluent 3ml×1 bottle 6ml×1 bottle 2-8℃
Chromogen Solution A 3ml×1 bottle 6ml×1 bottle 2-8℃
Chromogen Solution B 3ml×1 bottle 6ml×1 bottle 2-8℃
Stop Solution 3ml×1 bottle 6ml×1 bottle 2-8℃
wash solution (20ml×20 fold)
×1bottle (20ml×30 fold)
×1bottle 2-8℃
Specimen requirements
1.serum- coagulation at room temperature 10-20 mins,centrifugation 20-min at the speed of 2000-3000 r.p.m. remove supernatant, If precipitation appeared, Centrifugal again.
2.plasma-use suited EDTA or citrate plasma as an anticoagulant,mix 10-20 mins ,centrifugation 20-min at the speed of 2000-3000 r.p.m. remove supernatant, If precipitation appeared, Centrifugal again.
3.Urine-collect sue a sterile container, centrifugation 20-min at the speed of 2000-3000 r.p.m. remove supernatant, If precipitation appeared, Centrifugal again. The Operation of Hydrothorax and cerebrospinal fluid Reference to it.
4.cell culture supernatant-detect secretory components, collect sue a sterile container, centrifugation 20-min at the speed of 2000-3000 r.p.m. remove supernatant,detect the composition of cells, Dilut cell suspension with PBS(PH7.2-7.4), Cell concentration reached 1 million / ml, repeated freeze-thaw cycles, damage cells and release of intracellular components, centrifugation 20-min at the speed of 2000-3000 r.p.m. remove supernatant, If precipitation appeared, Centrifugal again.
5.Tissue samples- After cutting samples, check the weight,add PBS(PH7.2-7.4), Rapidly frozen with liquid nitrogen, maintain samples at 2-8℃ after melting,add PBS(PH7.4), Homogenized by hand or Grinders, centrifugation 20-min at the speed of 2000-3000 r.p.m. remove supernatant.
6.extract as soon as possible after Specimen collection,and according to the relevant literature, and should be experiment as soon as possible after the extraction. If it can’t, specimen can be kept in -20 ℃ to preserve, Avoid repeated freeze-thaw cycles.
7.Can’t detect the sample which contain NaN3, because NaN3 inhibits HRP active.
Assay procedure
1.Dilute and add sample to Standard: set 10 Standard wells on the ELISA plates coated, add Standard 100μl to the first and the second well, then add Standard dilution 50μl to the first and the second well, mix; take out 100μl form the first and the second well then add it to the third and the forth well separately. then add Standard dilution 50μl to the third and the forth well ,mix ; then take out 50μl from the third and the forth well discard, add 50μl to the fifth and the sixth well ,then add Standard dilution 50μl to the fifth and the sixth well, mix ; take out 50μl from the fifth and the sixth well and add to the seventh and the eighth well, then add Standard dilution 50μl to the seventh and the eighth well ,mix ; take out 50μl from the seventh and the eighth well and add to the ninth and the tenth well, add Standard dilution 50μl to the ninth and the tenth well, mix , take out 50μl from the ninth and the tenth well discard(add Sample 50μl to each well after Diluting ,(density: 240ng/L,160ng/L ,80ng/L,40ng/L, 20ng/L)
2.add sample:Set blank wells separately (blank comparison wells .don’t add sample and HRP-Conjugate reagent, other each step operation is same). testing sample well. add Sample dilution 40μl to testing sample well, then add testing sample 10μl (sample final dilution is 5-fold), add sample to wells , don’t touch the well wall as far as possible, and Gently mix.
3.Incubate: After closing plate with Closure plate membrane ,incubate for 30 min at 37℃.
4.Configurate liquid: 30-fold (or 20-fold)wash solution diluted 30-fold (or 20-fold) with distilled water and reserve.
5.washing:Uncover Closure plate membrane, discard Liquid, dry by swing, add washing buffer to every well, still for 30s then drain, repeat 5 times, dry by pat.
6.add enzyme:Add HRP-Conjugate reagent 50μl to each well, except blank well.
7.incubate:Operation with 3.
8.washing:Operation with 5.
9.color:Add Chromogen Solution A 50ul and Chromogen Solution B to each well, evade the light preservation for 15 min at 37℃
10.Stop the reaction:Add Stop Solution50μl to each well, Stop the reaction(the blue color change to yellow color).
11.assay:take blank well as zero , Read absorbance at 450nm after Adding Stop Solution and within 15min.
Important notes
1.The kit takes out from the refrigeration environment should be balanced 15-30 minutes in the room temperature, ELISA plates coated if has not use up after opened, the plate should be stored in Sealed bag.
2.washing buffer will Crystallization separation, it can be heated the water helps dissolve when dilute . Washing does not affect the result.
3.add Sample with sampler Each step, And proofread its accuracy frequently, avoids the experimental error. add sample within 5 mins, if the number of sample is much , recommend to use Volley .
4.if the testing material content is excessively higher (The sample OD is bigger than the first standard well ),please dilute Sample (n-fold), Please diluente and multiplied by the dilution factor.(×n×5).
5.Closure plate membrane only limits the disposable use, to avoid cross-contamination.
6.The substrate evade the light preservation.
7.Please according to use instruction strictly, The test result determination must take the microtiter plate reader as a standard.
8.All samples, washing buffer and each kind of reject should according to infective material process.
9.Do not mix reagents with those from other lots.
Calculate:
Take the standard density as the horizontal, the OD value for the vertical ,draw the standard curve on graph paper, Find out the corresponding density according to the sample OD value by the Sample curve, multiplied by the dilution multiple, or calculate the straight line regression equation of the standard curve with the standard density and the OD value ,with the sample OD value in the equation, calculate the sample density, multiplied by the dilution factor, the result is the sample actual density.
Storage and validity
1.Storage: 2-8℃.
2.validity: six months.
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Pierce SA510010 Anti-Rat IgM, Cross Absorbed 0.5 mg
Pierce SA510011 Anti-Rat IgM, Cross Absorbed 0.5 mg
Pierce SA510012 Anti-Rat IgM, Cross Absorbed 0.5 mg
Pierce SA510013 Anti-Rat IgM, Cross Absorbed 0.5 mg
Pierce SA510014 Anti-Rat IgM, Cross Absorbed 0.5 mg
Pierce SA510015 Anti-Rat IgM, Cross Absorbed 0.5 mg
Pierce SA510016 Anti-Rat IgM, Cross Absorbed 0.5 mg
Pierce SA510017 Anti-Rat IgG (H+L), Cross Absorbed 0.5 mg
Pierce SA510018 Anti-Rat IgG (H+L), Cross Absorbed 0.5 mg
Pierce SA510019 Anti-Rat IgG (H+L), Cross Absorbed 0.5 mg
Pierce SA510020 Anti-Rat IgG (H+L), Cross Absorbed 0.5 mg
Pierce SA510021 Anti-Rat IgG (H+L), Cross Absorbed 0.5 mg
Pierce SA510022 Anti-Rat IgG (H+L), Cross Absorbed 0.5 mg
Pierce SA510023 Anti-Rat IgG (H+L), Cross Absorbed 0.5 mg
Pierce SA510024 Anti-Rat IgG (H+L), Cross Absorbed 0.5 mg
Pierce SA510025 Anti-Rat IgG (H+L), Cross Absorbed 0.5 mg
Pierce SA510026 Anti-Rat IgG (H+L), Cross Absorbed 0.5 mg
Pierce SA510027 Anti-Rat IgG (H+L), Cross Absorbed 0.5 mg
Pierce SA510028 Anti-Rat IgG (H+L), Cross Absorbed 0.5 mg
Pierce SA510029 Anti-Rat IgG (H+L), Cross Absorbed 0.5 mg
Pierce SA510030 Anti-Rat IgG (H+L), Cross Absorbed 0.5 mg
Pierce SA510031 Anti-Rat IgG (H+L), Cross Absorbed 0.5 mg
Pierce SA510032 Anti-Rat IgG (H+L), Cross Absorbed 0.5 mg
Pierce SA510033 Anti-Rabbit IgG (H+L), Cross Absorbed 0.5 mg
Pierce SA510034 Anti-Rabbit IgG (H+L), Cross Absorbed 0.5 mg
Pierce SA510035 Anti-Rabbit IgG (H+L), Cross Absorbed 0.5 mg
Pierce SA510036 Anti-Rabbit IgG (H+L), Cross Absorbed 0.5 mg
Pierce SA510037 Anti-Rabbit IgG (H+L), Cross Absorbed 0.5 mg
Pierce SA510038 Anti-Rabbit IgG (H+L), Cross Absorbed 0.5 mg
Pierce SA510039 Anti-Rabbit IgG (H+L), Cross Absorbed 0.5 mg
Pierce SA510040 Anti-Rabbit IgG (H+L), Cross Absorbed 0.5 mg
Pierce SA510041 Anti-Rabbit IgG (H+L), Cross Absorbed 0.5 mg
Pierce SA510042 Anti-Rabbit IgG (H+L), Cross Absorbed 0.5 mg
Pierce SA510043 Anti-Rabbit IgG (H+L), Cross Absorbed 0.5 mg
Pierce SA510044 Anti-Rabbit IgG (H+L), Cross Absorbed 0.5 mg
Pierce SA510049 Anti-Rabbit IgG (H+L), Cross Absorbed 0.5 mg
Pierce SA510053 Anti-Sheep IgG (H+L), Cross Absorbed 0.5 mg
Pierce SA510054 Anti-Sheep IgG (H+L), Cross Absorbed 0.5 mg
Pierce SA510055 Anti-Sheep IgG (H+L), Cross Absorbed 0.5 mg
Pierce SA510056 Anti-Sheep IgG (H+L), Cross Absorbed 0.5 mg
Pierce SA510057 Anti-Sheep IgG (H+L), Cross Absorbed 0.5 mg
Pierce SA510058 Anti-Sheep IgG (H+L), Cross Absorbed 0.5 mg
Pierce SA510059 Anti-Sheep IgG (H+L), Cross Absorbed 0.5 mg
Pierce SA510060 Anti-Sheep IgG (H+L), Cross Absorbed 0.5 mg
Pierce SA510061 Anti-Donkey IgG (H+L) 0.5 mg
Pierce SA510062 Anti-Donkey IgG (H+L) 0.5 mg
Pierce SA510063 Anti-Donkey IgG (H+L) 0.5 mg
Pierce SA510064 Anti-Donkey IgG (H+L) 0.5 mg
Pierce SA510065 Anti-Donkey IgG (H+L) 0.5 mg
Pierce SA510066 Anti-Donkey IgG (H+L) 0.5 mg
Pierce SA510067 Anti-Donkey IgG (H+L) 0.5 mg
Pierce SA510068 Anti-Donkey IgG (H+L) 0.5 mg
Pierce SA510069 Anti-Chicken IgG (IgY) (H+L), Cross Absorbed 0.5 mg
Pierce SA510070 Anti-Chicken IgG (IgY) (H+L), Cross Absorbed 0.5 mg
Pierce SA510071 Anti-Chicken IgG (IgY) (H+L), Cross Absorbed 0.5 mg
Pierce SA510072 Anti-Chicken IgG (IgY) (H+L), Cross Absorbed 0.5 mg
Pierce SA510073 Anti-Chicken IgG (IgY) (H+L), Cross Absorbed 0.5 mg
Pierce SA510074 Anti-Chicken IgG (IgY) (H+L), Cross Absorbed 0.5 mg
Pierce SA510075 Anti-Chicken IgG (IgY) (H+L), Cross Absorbed 0.5 mg