ELISA 檢測的目的是為實(shí)驗(yàn)提供準(zhǔn)確可靠的定量分析實(shí)驗(yàn)依據(jù)。為了保證實(shí)驗(yàn)數(shù)據(jù)的可靠性，在實(shí)驗(yàn)過程中必須堅(jiān)持全面質(zhì)量控制和全過程質(zhì)量控制。在收集標(biāo)本前都必須有一個(gè)完整的計(jì)劃。齊一生物銷售：021－6034 8496；181214 53965；173021 04490

注意事項(xiàng)：

◆   每個(gè)標(biāo)本量收集體積＝  100ul × 檢測種類，如要做復(fù)孔，標(biāo)本量收集體積＝  100ul × 檢測種類×2 。

◆   對(duì)于作某一個(gè)具體指標(biāo)來說，先用一系列稀釋度作一批標(biāo)本測定，得出對(duì)實(shí)驗(yàn)有意義的一個(gè)稀釋度（即測定劑量反應(yīng)曲線），然后用一個(gè)*適稀釋度測定即可。

◆   收集標(biāo)本前必須清楚要檢測的成份是否足夠穩(wěn)定。對(duì)收集后當(dāng)天進(jìn)行檢測的標(biāo)本，儲(chǔ)存在4℃?zhèn)溆�，如有特殊原因需要周期收集�?biāo)本，將標(biāo)本及時(shí)分裝后放在-20℃ 或-70℃ 條件下保存。避免反復(fù)凍融。標(biāo)本 2 -8 ℃ 可保存48 小時(shí).-20℃ 可保存1個(gè)月。 -70 度可保存6個(gè)月。部分激素類標(biāo)本需添加抑肽酶。

◆   血清標(biāo)本采集時(shí)，應(yīng)注意避免溶血，紅細(xì)胞溶解時(shí)會(huì)釋放出具有過氧化物酶活性的物質(zhì)，以 HRP為標(biāo)記的 ELISA 測定中，溶血標(biāo)本可能會(huì)增加非特異性顯色。

【小鼠B細(xì)胞淋巴瘤因子2(Bcl-2)ELISA試劑盒】◆   為了保證尿液檢測結(jié)果的準(zhǔn)確性，必須正確收集尿液標(biāo)本和保存。盛尿容器要清潔干燥。使用一次性的容器（如塑料尿杯），避免因用藥并清洗不干凈而造成的污染，影響檢測結(jié)果。尿液標(biāo)本必須新鮮，留取后，應(yīng)及時(shí)檢測或保存，以免細(xì)菌繁殖。因在室溫中久置后（尤其是夏季）尿中磷酸鹽等可析出結(jié)晶而干擾檢測。

◆    標(biāo)本宜在新鮮時(shí)檢測。如有細(xì)菌污染，菌體中可能含有內(nèi)源性 HRP，也會(huì)產(chǎn)生假陽性反應(yīng)。保存過久可發(fā)生聚合在 ELISA中可使本底加深。

◆   凍結(jié)標(biāo)本融解后，蛋白質(zhì)局部濃縮，分布不均，應(yīng)充分混勻宜輕緩，避免氣泡，可上下顛倒混和，不要在混勻器上強(qiáng)烈振蕩。

◆   混濁或有沉淀的標(biāo)本應(yīng)先離心或過濾，澄清后再檢測。

◆   反復(fù)凍融會(huì)使蛋白效價(jià)降低，所以待測標(biāo)本如需保存作多次檢測，宜少量分裝冰存。也可加入適當(dāng)防腐劑。

◆   如果您在實(shí)驗(yàn)中遇到任何特殊問題，敬請(qǐng)咨詢 021-60348496標(biāo)本類型

ELISA常見的標(biāo)本一般包括：

◆   液體類標(biāo)本：包括血清、血漿、尿液、胸腹水、腦脊液、細(xì)胞培養(yǎng)上清等。

◆   培養(yǎng)細(xì)胞

◆   組織標(biāo)本

這些標(biāo)本收集的時(shí)間、方法和保存都有一定的要求。

【小鼠B細(xì)胞淋巴瘤因子2(Bcl-2)ELISA試劑盒】標(biāo)本的保存

一般說來，在 5 天內(nèi)測定的標(biāo)本可放置于 4℃，超過一周測定的需低溫凍存。

對(duì)收集后當(dāng)天進(jìn)行檢測的標(biāo)本，儲(chǔ)存在  4 ℃ 備用，如有特殊原因需要周期收集標(biāo)本， 將標(biāo)本及時(shí)分裝后放在  -20 ℃ 或  -70 ℃ 條件下保存。避免反復(fù)凍融。 標(biāo)本  2 -8 ℃ 可保存  48 小時(shí)，-20 ℃ 可保存  1 個(gè)月。-70 度可保存  6 個(gè)月。部分標(biāo)本也可加入適當(dāng)防腐劑，激素類標(biāo)本需添加抑肽酶。標(biāo)本的處理 可用作 ELISA 測定的標(biāo)本十分廣泛，體液（如血清）、分泌物（唾液）和排泄物（如尿液）、細(xì)胞培養(yǎng)上清、細(xì)胞、組織等均可作標(biāo)本以測定其中某種成份。有些標(biāo)本可直接進(jìn)行測定，有些則需經(jīng)預(yù)處理。

◆ 血清：  

室溫血液自然凝固  10-20 分鐘后，離心  20 分鐘左右（  2000-3000 轉(zhuǎn)  / 分）。收集上清。如有沉淀形成，應(yīng)再次離心。  

◆ 血漿：  

應(yīng)根據(jù)試劑盒的要求選擇  EDTA 、檸檬酸鈉或肝素作為抗凝劑，加入  10 ％（  v/v ）抗凝劑 ( 0.1M 檸檬酸鈉或  1% heparin 或  2.0%EDTA.Na2) 混合  10-20 分鐘后，離心  20 分鐘左右（  2000-3000 轉(zhuǎn)  / 分）。仔細(xì)收集上清如有沉淀形 成，應(yīng)再次離心。  

【小鼠B細(xì)胞淋巴瘤因子2(Bcl-2)ELISA試劑盒】◆ 尿液、胸腹水、腦脊液：  

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。如有沉淀形成，應(yīng)再次離心。  

◆ 細(xì)胞培養(yǎng)上清：  

檢測分泌性的成份時(shí)，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(shí)，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。  

◆ 細(xì)胞：

檢測細(xì)胞的成份時(shí)，對(duì)于貼壁細(xì)胞，去除培養(yǎng)液，用PBS、生理鹽水或無血清培養(yǎng)液洗一遍。加入適量裂解液。用槍吹打數(shù)下，使裂解液和細(xì)胞充分接觸。通常裂解液接觸細(xì)胞10 秒后，細(xì)胞就會(huì)被裂解。對(duì)于懸浮細(xì)胞，離心收集細(xì)胞，用PBS、生理鹽水或無血清培養(yǎng)液洗一遍.加入適量裂解液, 用槍吹打把細(xì)胞吹散。用手指輕彈以充分裂解細(xì)胞。充分裂解后應(yīng)沒有明顯的細(xì)胞沉淀。如果細(xì)胞量較多，必需分裝然后再裂解。充分裂解后，10000-14000g離心3-5 分鐘，取上

清，即可進(jìn)行后續(xù)操作。

【小鼠B細(xì)胞淋巴瘤因子2(Bcl-2)ELISA試劑盒】◆ 組織標(biāo)本：  

切割標(biāo)本后，稱取重量。加入一定量的  PBS或組織蛋白萃取試劑，緩沖液中可加入蛋白酶抑制劑。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)收集上清置于  -20 度或  - 70 度保存，如有必要，可以將樣品濃縮干燥。分裝后一份待檢測，其余冷凍備用。齊一生物銷售：021－6034 8496；181214 53965；173021 04490  

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