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首頁 > 蛋白生物學 > 蛋白Marker > 多肽電泳Marker

超低分子量蛋白Marker I(3.3-22KD)

超低分子量蛋白Marker I(3.3-22KD)

產品(pin)編號:RTD6101

產品規格:10次

數量
價格 ¥200

超(chao)低分子量蛋白質(zhi)Marker I3.3-22kD

Ultra Low Molecular Weight Protein Marker I

產品編號及規格:

RTD6101   10T100μl)   200元

 儲存條件(jian):

   -20保存,開封(feng)后有效期12個月。

 產品簡介:

本產品包含3種多肽(tai)和(he)2種(zhong)低(di)分(fen)子量蛋白質組成,分(fen)子量范圍為3.3kD-22kD。可以(yi)用來判(pan)斷 SDS-PAGE上多肽和小蛋白的分(fen)子量(liang)。


● 使(shi)用說明:

第一次收到該產品,常溫融化后,徹底混勻,離心快甩將溶液完全收集到管底

. 制膠:

I 配制分離膠

1.按照表一將不同體積的雙蒸水、40% PAA19:1)、4×凝膠緩沖液(貨號:GB010)和乙二醇加入到小燒杯中混(hun)合。

2.加入10%APSTEMED,立即混勻5-10秒,以(yi)使溶(rong)液(ye)充分(fen)混(hun)勻。

3.在凝膠模具中迅速灌入適量分離膠溶液,然后在分離膠溶液上輕輕覆蓋一層1 cm的無水乙醇,使凝膠表面保持(chi)平整(zheng)。

4.靜置15-30分鐘,待分離膠和乙醇(chun)層之間出現一個清晰(xi)的界(jie)面表(biao)示凝膠已聚合。

表一  (一塊1 mm mini膠用量)

 

分離(li)膠

濃(nong)縮膠

 

18T, 5%C /5 ml

5T, 3.3%C/2 ml

40% PAA19:1

2.25 ml

/

40% PAA (29:1)

/

0.25 ml

4×凝膠緩沖液

1.25 ml

0.5 ml

乙二醇(電泳級)

1.5 ml

/

ddH2O

/

1.25ml

10APS

50 μl

20 μl

TEMED

5 μl

2 μl


II 配制濃縮膠

去除覆(fu)蓋在分離膠上的乙醇層,用濾紙將(jiang)殘留乙醇吸去。

1.按(an)照(zhao)表一將不同體(ti)積的雙蒸水(shui)、40%PAA(29:1)和4×凝膠緩沖液加(jia)入到小(xiao)燒杯中混合。

2.加入10%過硫(liu)酸銨(an)和(he)TEMED,立即混勻5-10秒,以使溶(rong)液充分混勻。

3.將梳子插入凝膠內(nei),避免產生氣(qi)泡(pao)。

4.靜置30-60分(fen)鐘裝待(dai)凝膠聚合

. 電泳:

1. Marker樣品,充分混勻后,95℃處理5分鐘。

2. 電泳(yong)前,將10×陽極(ji)(ji)緩(huan)(huan)(huan)沖(chong)(chong)(chong)液(Cat No:AB080)和10×陰(yin)極(ji)(ji)緩(huan)(huan)(huan)沖(chong)(chong)(chong)液(Cat No:CB010)用(yong)蒸(zheng)餾水稀釋(shi)成(cheng)1×緩(huan)(huan)(huan)沖(chong)(chong)(chong)液備用(yong)。將電泳(yong)槽(cao)(cao)的外(wai)槽(cao)(cao)加(jia)入(ru)1×陽極(ji)(ji)緩(huan)(huan)(huan)沖(chong)(chong)(chong)液,內槽(cao)(cao)加(jia)入(ru)1×陰(yin)極(ji)(ji)緩(huan)(huan)(huan)沖(chong)(chong)(chong)液,輕柔拔出梳子(zi),將Marker或(huo)蛋白樣品(已經過2×Tricine上樣緩(huan)(huan)(huan)沖(chong)(chong)(chong)液[Cat No:TP050]處理)加(jia)入(ru)點樣孔(kong),參考下表(biao)進行(xing)電泳(yong),至每條帶都清晰可見時停止電泳(yong)。整個(ge)電泳(yong)過程大(da)約需(xu)要2-3個(ge)小時。

表二 多肽電泳條件

恒電壓

150V

起始電流

60-75mA/板膠

結束電流

15-25mA/板(ban)膠

電泳時間(jian)

2.5-3小時




多肽電泳配套試劑

 


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