常温化学镀锌配方纯化军绿色用哪种配方好

来源：蜘蛛抓取(WebSpider) 时间：2016-12-28 19:54 标签： 化学镀锌的配方
supplierchannel镀锌纯化DZn8是什么意思?
【圣血三爷】85
镀锌纯化DZn8是一般垫圈的标注,DZn----表示镀锌,8-----表示垫圈的规格为 8.标注齐全适应注明国标号,如 D.Zn8/GB/T95-1985.
不是垫圈是有外螺纹的直角过渡接头！
直角过渡接头的标注，DZn镀锌是对的，DZn8标注不符合国标，执行标准是否企标。
为您推荐：
其他类似问题
扫描下载二维码关注今日：1 | 主题：387471
微信扫一扫
扫一扫，下载丁香园 App
即送15丁当
我该用哪种ＧＳＴ融合蛋白纯化试剂盒
页码直达：
这个帖子发布于11年零353天前，其中的信息可能已发生改变或有所发展。
我用的是ＰＧＥＸ－６Ｐ－１质粒，已经表达，请问我用哪种纯化试剂盒合适？有没有相关信息？我们这里做分子生物学的特少，信息不通畅
不知道邀请谁？试试他们
微信扫一扫
广告宣传推广
政治敏感、违法虚假信息
恶意灌水、重复发帖
违规侵权、站友争执
附件异常、链接失效
一般不需要纯化试剂盒，买点带Glutathine sepharose即可，Amersham和Promega就有卖的，操作步骤他们网站上有。
微信扫一扫
广告宣传推广
政治敏感、违法虚假信息
恶意灌水、重复发帖
违规侵权、站友争执
附件异常、链接失效
微信扫一扫
广告宣传推广
政治敏感、违法虚假信息
恶意灌水、重复发帖
违规侵权、站友争执
附件异常、链接失效
我用的是国外的一位作者给邮过来的ｐGEX－载体质粒，已经表达，我想纯化一下，然后用纯化的蛋白作为我的激酶反应的一个底物，请问该如何纯化，用试剂盒吗？还是直接买珠子就行啊？并且用哪个公司的好啊，价钱还合适！谢谢各位了先。
微信扫一扫
广告宣传推广
政治敏感、违法虚假信息
恶意灌水、重复发帖
违规侵权、站友争执
附件异常、链接失效
你不需要Kits,按这个方法做,100%成功Purification of GST Fused Proteins Author: Chia Jin Ngee Source: Contributed by Chia Jin Ngee Abstract: Many people have vented out frustration over insoluble GST-fused proteins. This is a protocol for enzymatically active soluble GST-fused proteins. All GST-fused proteins are rendered soluble with this technique though enzyme activitiy can range from 30-90%.
-------------------------------------------------------------------------------- Materials and Reagents STE Buffer 10 mM Tris-HCl, pH 8.01 mM EDTA150 mM NaCl Lysozyme solution 10 mg/ml in water (make fresh) PBS Elution Buffer 50 mM Tris.Cl, pH 9.020 mM GSH 10% Sarkosyl in STE Buffer 10% Triton X-100 in STE Buffer 1 M DTT 100 mM IPTG Procedure
Day 1 Set up an overnight culture in 50 ml 2XTY with 150 mg/ml of ampicillin.
Day 2 Seed 5 ml of overnight culture to 500 ml 2XTY with 150 mg/ml of ampicillin. Grow at 37oC to an A600 of 0.6 to 0.8. Induce with 0.1 mM to 2 mM of IPTG. Grow for 3 hr at 37oC or grow overnight at room temperature.Lower IPTG concentrations and lower growing temperatures tend to produce greater solubility at the expense of yield. Pellet cells by centrifuging at 3000 g, 4oC for 10 min. Decant media and resuspend cells in 30 ml ice-cold PBS to wash. Transfer to a 40-ml Oak Ridge tube and centrifuge at 3000 g, 4oC for 10 min. Decant PBS. This is a convenient point to stop and to store pellets at -80oC. Else continue to lyse cells. Thaw pellet on ice if cells are frozen else proceed to the next step. Resuspend pellet in 10 ml of ice cold STE Buffer. Add 100 ml of freshly prepared lyozyme solution, incubate on ice for 15 min. Just before sonication, add 100 ml of 1 M DTT and 1.4 ml of 10% Sarkosyl. Mix thoroughly by inversion and sonicate for a total time of 1 min. Centrifuge 16,000 rpm
for 20 min on the SS34 rotor to pellet debris. Transfer supernatant to a 50-ml conical tube and discard the pellet. Add 4 ml of 10% Triton X-100 and top up with STE Buffer to 20 ml. The effective concentration of Sarkosyl and Triton X-100 will be 0.7% and 2% respectively. Incubate at room temperature for 30 min. Pour the lysate to 1 ml bed of prepared Glutathione Sepharose in PBS. Incubate at room temperature for 30 min to 1 hr with agitation.To prepare the 50% slurry, shake up the media and pipette 2 ml to a 50 ml tube. Fill to 50 ml with PBS, invert tube a few times. Centrifuge to 2000 rpm on a swing bucket centrifuge then switch off. Carefully suck off PBS and resuspend beads with 1 ml of PBS. Wash the beads with 3 X 50 ml of PBS. Finally resuspend in 5 ml of PBS. Pour to a dispo-column. Wash the 50-ml conical tube with an additional 5 ml of PBS. Pool with the first 5 ml in the dispo-column. To wash, use the same centrifugation technique for preparing the beads. When transferring beads to column, do not pipette but pour. The beads tend to stick to pipette tips. If desired, elute with 10 x 1 ml fractions of Elution Buffer. Determine desired fractions with SDS PAGE. Reference Frangioni and Neel.Anal. Biochem. 210, 179-187 (1993)
微信扫一扫
广告宣传推广
政治敏感、违法虚假信息
恶意灌水、重复发帖
违规侵权、站友争执
附件异常、链接失效
融合蛋白是包涵体么？您最后纯化成功了么？用什么方法呢？
微信扫一扫
广告宣传推广
政治敏感、违法虚假信息
恶意灌水、重复发帖
违规侵权、站友争执
附件异常、链接失效
关于丁香园

我要回帖

更多关于 化学镀锌的配方 的文章

 

随机推荐