人白細(xì)胞抗原E(HLA-E)酶聯(lián)免疫分析(ELISA)試劑盒用于測定人血清,血漿及相關(guān)液體樣本中白細(xì)胞抗原E(HLA-E)的含量。應(yīng)用雙抗體夾心法測定標(biāo)本中人白細(xì)胞抗原E(HLA-E)水平。 試劑盒組成: 試劑盒組成 | 48孔配置 | 96孔配置 | 保存 | 說明書 | 1份 | 1份 | | 封板膜 | 2片(48) | 2片(96) | | 密封袋 | 1個 | 1個 | | 酶標(biāo)包被板 | 1×48 | 1×96 | 2-8℃保存 | 標(biāo)準(zhǔn)品:13.5U/ml | 0.5ml×1瓶 | 0.5ml×1瓶 | 2-8℃保存 | 標(biāo)準(zhǔn)品稀釋液 | 1.5ml×1瓶 | 1.5ml×1瓶 | 2-8℃保存 | 酶標(biāo)試劑 | 3 ml×1瓶 | 6 ml×1瓶 | 2-8℃保存 | 樣品稀釋液 | 3 ml×1瓶 | 6 ml×1瓶 | 2-8℃保存 | 顯色劑A液 | 3 ml×1瓶 | 6 ml×1瓶 | 2-8℃保存 | 顯色劑B液 | 3 ml×1瓶 | 6 ml×1瓶 | 2-8℃保存 | 終止液 | 3ml×1瓶 | 6ml×1瓶 | 2-8℃保存 | 濃縮洗滌液 | (20ml×20倍)×1瓶 | (20ml×30倍)×1瓶 | 2-8℃保存 |
樣本處理及要求: 1. 血清:室溫血液自然凝固10-20分鐘,離心20分鐘左右(2000-3000轉(zhuǎn)/分)。仔細(xì)收集上清,保存過程中如出現(xiàn)沉淀,應(yīng)再次離心。 2. 血漿:應(yīng)根據(jù)標(biāo)本的要求選擇EDTA或檸檬酸鈉作為抗凝劑,混合10-20分鐘后,離心20分鐘左右(2000-3000轉(zhuǎn)/分)。仔細(xì)收集上清,保存過程中如有沉淀形成,應(yīng)該再次離心。 3. 尿液:用無菌管收集,離心20分鐘左右(2000-3000轉(zhuǎn)/分)。仔細(xì)收集上清,保存過程中如有沉淀形成,應(yīng)再次離心。胸腹水、腦脊液參照實行。 4. 細(xì)胞培養(yǎng)上清:檢測分泌性的成份時,用無菌管收集。離心20分鐘左右(2000-3000轉(zhuǎn)/分)。仔細(xì)收集上清。檢測細(xì)胞內(nèi)的成份時,用PBS(PH7.2-7.4)稀釋細(xì)胞懸液,細(xì)胞濃度達(dá)到100萬/ml左右。通過反復(fù)凍融,以使細(xì)胞破壞并放出細(xì)胞內(nèi)成份。離心20分鐘左右(2000-3000轉(zhuǎn)/分)。仔細(xì)收集上清。保存過程中如有沉淀形成,應(yīng)再次離心。 5. 組織標(biāo)本:切割標(biāo)本后,稱取重量。加入一定量的PBS,PH7.4。用液氮迅速冷凍保存?zhèn)溆谩?biāo)本融化后仍然保持2-8℃的溫度。加入一定量的PBS(PH7.4),用手工或勻漿器將標(biāo)本勻漿充分。離心20分鐘左右(2000-3000轉(zhuǎn)/分)。仔細(xì)收集上清。分裝后一份待檢測,其余冷凍備用。 6. 標(biāo)本采集后盡早進(jìn)行提取,提取按相關(guān)文獻(xiàn)進(jìn)行,提取后應(yīng)盡快進(jìn)行實驗。若不能馬上進(jìn)行試驗,可將標(biāo)本放于-20℃保存,但應(yīng)避免反復(fù)凍融. 7. 不能檢測含NaN3的樣品,因NaN3抑制辣根過氧化物酶的(HRP)活性。 試劑盒性能: 1.樣品線性回歸與預(yù)期濃度相關(guān)系數(shù)R值為0.92以上。 2.批內(nèi)與批見應(yīng)分別小于9%和15% 檢測范圍: 0.15U/ml -9U/ml 1.試劑盒保存:2-8℃ 2.有效期:6個月 注:僅供科學(xué)研究實驗使用,內(nèi)容供參考,具體信息請咨詢。 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. |