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小鼠中性粒細胞明膠酶相關(guān)脂質(zhì)運載蛋白酶免試劑盒,(NGAL)ELISA檢測試劑盒 ,英文名:NGAL ELISA Kit ,英文縮寫:NGAL ,常見試劑盒規(guī)格:96T/48T,產(chǎn)品別名:小鼠中性粒細胞明膠酶相關(guān)脂質(zhì)運載蛋白ELISA檢測試劑盒,NGAL 檢測試劑盒, NGAL 檢測試劑盒
ELISA實驗操作較繁雜,操作不當(dāng)將引起較大的誤差。小鼠中性粒細胞明膠酶相關(guān)脂質(zhì)運載蛋白酶免試劑盒,(NGAL)ELISA檢測試劑盒在操作中應(yīng)注意以下5個方面。
1.隨著病情的進展或由其他因素影響,某些抗體在機體內(nèi)的含量發(fā)生大幅波動,因此有必要對標本進行預(yù)處理,例如對血清標本作適當(dāng)稀釋,或離心分離血脂等。間接法測定中,標本適當(dāng)稀釋還可降低非特異性反應(yīng)。
2.加樣一般使用加液器,在ELISA中一般有4次加樣:加標本、加酶結(jié)合物、加底物和加終止液。加標本時必須每份標本使用一支移液器吸嘴,以免交叉污染。加酶結(jié)合物、底物和終止液時則不需更換吸嘴。
3.在成批手工檢測中,還應(yīng)注意操作時差的影響。加樣及混勻時間的差異,使抗原抗體反應(yīng)和酶促反應(yīng)時間不*,對競爭法及定量檢測影響很大,不注意可能會導(dǎo)致錯誤結(jié)果或定量不準。
4.洗滌是ELISA操作過程中決定實驗成敗的一個關(guān)鍵步驟。目的是除去未結(jié)合的免疫反應(yīng)物,終止抗原抗體反應(yīng),除去標本中與反應(yīng)無關(guān)的成分、游離的酶標物以及反應(yīng)過程中吸附于固相載體上的非特異性物質(zhì)??捎孟窗鍣C洗板或手工洗板,前者洗滌質(zhì)量較好,各反應(yīng)孔洗滌效果均一,批內(nèi)、批間差異小,手工洗板應(yīng)注意控制各孔洗滌效果,差異不宜太大。
5.準確判定結(jié)果須使用ELISA測讀儀(酶標儀),比色法判讀可選擇單波長或雙波長,根據(jù)反應(yīng)液顏色的不同選用不同波長的濾光片,以空白孔校零測定反應(yīng)孔的吸光度值(A值)。酶標儀具有良好的重復(fù)性。
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各種類型ELISA測定時,一般需經(jīng)過以下步驟:固相包被→加待測樣品→溫育→洗滌+加酶標物→溫育→洗滌→加底物→溫育一加終止液→比色→判定結(jié)果。