人白细胞抗原E(HLA-E)酶联免疫分析(ELISA)试剂盒用于测定人血清,血浆及相关液体样本中白细胞抗原E(HLA-E)的含量。应用双抗体夹心法测定标本中人白细胞抗原E(HLA-E)水平。 试剂盒组成: 试剂盒组成 | 48孔配置 | 96孔配置 | 保存 | 说明书 | 1份 | 1份 | | 封板膜 | 2片(48) | 2片(96) | | 密封袋 | 1个 | 1个 | | 酶标包被板 | 1×48 | 1×96 | 2-8℃保存 | 标准品:13.5U/ml | 0.5ml×1瓶 | 0.5ml×1瓶 | 2-8℃保存 | 标准品稀释液 | 1.5ml×1瓶 | 1.5ml×1瓶 | 2-8℃保存 | 酶标试剂 | 3 ml×1瓶 | 6 ml×1瓶 | 2-8℃保存 | 样品稀释液 | 3 ml×1瓶 | 6 ml×1瓶 | 2-8℃保存 | 显色剂A液 | 3 ml×1瓶 | 6 ml×1瓶 | 2-8℃保存 | 显色剂B液 | 3 ml×1瓶 | 6 ml×1瓶 | 2-8℃保存 | 终止液 | 3ml×1瓶 | 6ml×1瓶 | 2-8℃保存 | 浓缩洗涤液 | (20ml×20倍)×1瓶 | (20ml×30倍)×1瓶 | 2-8℃保存 |
样本处理及要求: 1. 血清:室温血液自然凝固10-20分钟,离心20分钟左右(2000-3000转/分)。仔细收集上清,保存过程中如出现沉淀,应再次离心。 2. 血浆:应根据标本的要求选择EDTA或柠檬酸钠作为抗凝剂,混合10-20分钟后,离心20分钟左右(2000-3000转/分)。仔细收集上清,保存过程中如有沉淀形成,应该再次离心。 3. 尿液:用无菌管收集,离心20分钟左右(2000-3000转/分)。仔细收集上清,保存过程中如有沉淀形成,应再次离心。胸腹水、脑脊液参照实行。 4. 细胞培养上清:检测分泌性的成份时,用无菌管收集。离心20分钟左右(2000-3000转/分)。仔细收集上清。检测细胞内的成份时,用PBS(PH7.2-7.4)稀释细胞悬液,细胞浓度达到100万/ml左右。通过反复冻融,以使细胞破坏并放出细胞内成份。离心20分钟左右(2000-3000转/分)。仔细收集上清。保存过程中如有沉淀形成,应再次离心。 5. 组织标本:切割标本后,称取重量。加入一定量的PBS,PH7.4。用液氮迅速冷冻保存备用。标本融化后仍然保持2-8℃的温度。加入一定量的PBS(PH7.4),用手工或匀浆器将标本匀浆充分。离心20分钟左右(2000-3000转/分)。仔细收集上清。分装后一份待检测,其余冷冻备用。 6. 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. 7. 不能检测含NaN3的样品,因NaN3抑制辣根过氧化物酶的(HRP)活性。 试剂盒性能: 1.样品线性回归与预期浓度相关系数R值为0.92以上。 2.批内与批见应分别小于9%和15% 检测范围: 0.15U/ml -9U/ml 1.试剂盒保存:2-8℃ 2.有效期:6个月 注:仅供科学研究实验使用,内容供参考,具体信息请咨询。 Human leucocyte antigen-E Drug Names Generic Name:Human leucocyte antigen-E (HLA-E) ELISA Kit. Purpose This kit allows for the determination of HLA-Econcentrations in Human serum, plasma, and other biological fluids. Principle of the assay The kit assay Human HLA-Elevel in the sample,use Purified Human HLA-E antibody to coat microtiter plate wells, make solid-phase antibody, then add HLA-E to wells, Combined HLA-Eantibody which With HRP labeled , become antibody - antigen - enzyme-antibody complex, after washing Compley, Add TMB substrate solution,TMB substrate becomes blue color At HRP enzyme-catalyzed, reaction is terminated by the addition of a sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm. The concentration of HLA-E in the samples is then determined by comparing the O.D. of the samples to the standard curve. 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 range 0.15U/ml -9U/ml 1.Storage: 2-8℃. 2.validity: six montsh. |