产品名称:“猪白介素10 ELISA试剂盒 ” 本试剂盒用于体外定量检测血清、血浆、组织、细胞上清及相关液体样本中猪白细胞介素10(IL-10)的含量。 有效期:6个月 保存条件:2-8℃ 本试剂盒仅供体外研究使用,不用于临床诊断 实验原理 试剂盒采用双抗体一步夹心法酶联免疫吸附试验(ELISA)。往预先包被猪白细胞介素10(IL-10)捕获抗体的包被微孔中,依次加入标本、标准品、HRP标记的检测抗体,经过温育并彻底洗涤。用底物TMB显色,TMB在过氧化物酶的催化下转化成蓝色,并在酸的作用下转化成最终的黄色。颜色的深浅和样品中的猪白细胞介素10(IL-10)呈正相关。用酶标仪在450nm 波长下测定吸光度(OD 值),计算样品浓度。 试剂盒组成 130倍浓缩洗涤液20ml×1瓶7终止液6ml×1瓶 2酶标试剂6ml×1瓶8标准品(2400μg/L)0.5ml×1瓶 3酶标包被板12孔×8条9标准品稀释液1.5ml×1瓶 4样品稀释液6ml×1瓶10说明书1份 5显色剂A液6ml×1瓶11封板膜2张 6显色剂B液6ml×1/瓶12密封袋1个 标本要求 1.标本采集后尽早进行提取,提取按相关文献进行,提取后应尽快进行实验。若不能马上进行试验,可将标本放于-20℃保存,但应避免反复冻融 2.不能检测含NaN3的样品,因NaN3抑制辣根过氧化物酶的(HRP)活性。 “猪白介素10 ELISA试剂盒” 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。 1200μg/L5号标准品150μl的原倍标准品加入150μl标准品稀释液 600μg/L4号标准品150μl的5号标准品加入150μl标准品稀释液 300μg/L3号标准品150μl的4号标准品加入150μl标准品稀释液 150μg/L2号标准品150μl的3号标准品加入150μl标准品稀释液 75μg/L1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(样品最终稀释度为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30倍浓缩洗涤液用蒸馏水30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。 Materials provided with the kit 1wash solution20ml×1bottle7Stopp Solution6ml×1 bottle 2HRP-Conjugate reagent6ml×1 bottle8Standard(2400μg/L)0.5ml×1 bottle 3Microelisa stripplate12well×8strips9Standard diluent1.5ml×1bottle 4Sample diluent6ml×1 bottle10Instruction1 5Chromogen Solution A6ml×1 bottle11Closure plate membrane2 6Chromogen Solution B6ml×1 bottle12Sealed bags1 Assay procedure 1.Dilute and add sample:Dilute Original density Standard as follow table: 1200μg/L5 Standard150μl Original density Standard+150μl Standard diluent 600μg/L4 Standard150μl 5 Standard+150μl Standard diluent 300μg/L3 Standard150μl 4 Standard+150μl Standard diluent 150μg/L2 Standard150μl 3 Standard +150μl Standard diluent 75μg/L1 Standard150μl 2 Standard +150μl Standard diluent 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. “猪白介素10 ELISA试剂盒”价格/说明书欢迎来电索取,此产品本司提供免费代测,在接到客户标本当日起,现货产品一周内将检测报告交到客户手中!多年专业酶免服务,质量保证,期待您的来电!