小鼠内脂素/内脏脂肪素(visfatin)ELISA试剂盒含税含运费 , Elisa试剂盒价格,Elisa试剂盒说明书,Elisa试剂盒技术,Elisa试剂盒售后,Elisa试剂盒免费代测详情咨询: 电 话: 手 机: 传 真: ju111net手机版_ju111net登录_ju111.net登录导航长期提供各种种属、各种系列ELISA检测试剂盒,价格实惠,质量有保证,信誉*。索取各ELISA试剂盒产品说明书。凡购买目录本企业ELISA检测试剂盒可免费提供代测服务。本企业的更多产品,请点击企业:www.shhykit.com 产品规格:96T/48T 96T指可以做94个标本,2个标准对照 48T指可以做47个标本,1个标准对照 96T-84个样本 48T-42个样本 操作步骤 1.标准品的稀释:本试剂盒提供原倍标准品一支,用户可按照下列图表在小试管中进行稀释。 3600 pg/ml5号标准品150μl的原倍标准品加入150μl标准品稀释液 1800pg/ml4号标准品150μl的5号标准品加入150μl标准品稀释液 900pg/ml3号标准品150μl的4号标准品加入150μl标准品稀释液 450 pg/ml2号标准品150μl的3号标准品加入150μl标准品稀释液 225pg/ml1号标准品150μl的2号标准品加入150μl标准品稀释液 2.加样:分别设空白孔(空白对照孔不加样品及酶标试剂,其余各步操作相同)、标准孔、待测样品孔。在酶标包被板上标准品准确加样50μl,待测样品孔中先加样品稀释液40μl,然后再加待测样品10μl(样品zui终稀释度为5倍)。加样将样品加于酶标板孔底部,尽量不触及孔壁,轻轻晃动混匀。 3.温育:用封板膜封板后置37℃温育30分钟。 4.配液:将20倍浓缩洗涤液用蒸馏水20倍稀释后备用 5.洗涤:小心揭掉封板膜,弃去液体,甩干,每孔加满洗涤液,静置30秒后弃去,如此重复5次,拍干。 6.加酶:每孔加入酶标试剂50μl,空白孔除外。 7.温育:操作同3。 8.洗涤:操作同5。 9.显色:每孔先加入显色剂A50μl,再加入显色剂B50μl,轻轻震荡混匀,37℃避光显色15分钟. 10.终止:每孔加终止液50μl,终止反应(此时蓝色立转黄色)。 11.测定:以空白空调零,450nm波长依序测量各孔的吸光度(OD值)。 测定应在加终止液后15分钟以内进行。 Assay procedure 1.Dilute and add sample:Dilute Original density Standard as follow table: 800 nmol/L5 Standard150μl Original density Standard+150μl Standard diluent 400 nmol/L4 Standard150μl 5 Standard+150μl Standard diluent 200 nmol/L3 Standard150μl 4 Standard+150μl Standard diluent 100 nmol/L2 Standard150μl 3 Standard +150μl Standard diluent 50 nmol/L1 Standard150μl 2 Standard +150μl Standard diluent 2.add sample:Set blank wells separay (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 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. 小鼠内脂素/内脏脂肪素(visfatin)ELISA试剂盒免费代测,本试剂盒详细说明书欢迎您来电索取。 |