| 查看: 4842 | 回复: 12 | |||
果果gxl2013金虫 (小有名气)
|
[交流]
HRP与Peroxidase有区别吗？
|
||
各位，请问有人知道HRP与Peroxidase有区别吗？新买的二抗写着：Peroxidase-conjugated affinipure goat anti-rat IgG(H+L)；我买的一抗官网说明写着：Rat Anti-Mouse LDL R Monoclonal Antibody (Catalog # MAB2255) followed by HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005)。我做的从没出过条带，但是有背景。求大神解答 |
回复此楼» 猜你喜欢
莫那利珠单抗阻断NKG2A/HLA-E抑制通路 | Biochempartner
已经有0人回复
-
GSPT1口服分子胶降解剂MRT-2359 | Biochempartner
已经有0人回复
-
化学工程及工业化学论文润色/翻译怎么收费?
已经有204人回复
-
碳青霉烯类抗生素为何需联用西司他丁钠？ | Biochempartner
已经有0人回复
-
源自蕨类植物的抗炎美白活性分子去甲氧基荚果蕨素 | Biochempartner
已经有0人回复
-
肠道里的"代谢开关"：Fexaramine用肠道限制性重新定义 FXR 激动剂
已经有0人回复
-
DMXAA：从血管破坏剂到STING通路研究的关键工具分子 | Biochempartner
已经有0人回复
-
靶向PD-L1的人源化单克隆抗体阿特珠单抗
已经有0人回复
-
"探针之父"(+)-JQ-1如何改变表观遗传研究 | Biochempartner
已经有0人回复
-
含氟糖皮质激素的外用抗炎利器醋酸氟轻松
已经有0人回复
-
阿莫奈韦（Amenamevir）：解旋酶-引物酶靶向分子的结构特征与合成路径全解析
已经有0人回复
2楼2017-05-26 13:41:04
★ ★
小木虫: 金币+0.5, 给个红包，谢谢回帖
果果gxl2013: 金币+1 2017-06-03 13:37:23
小木虫: 金币+0.5, 给个红包，谢谢回帖
果果gxl2013: 金币+1 2017-06-03 13:37:23
|
????, ?????????^??????~ ????? SDS loading buffer ?c DTT ??????I??~ ????DTT ???????? DTT 250mM or 2-mercaptoethanol(?????F???е??) 12.5% (v/v) /5X sds loading buffer https://www.scientistsolutions.c ... mples#opennewwindow, ??????: Hello all, A co-worker of mine and I are debating the most efficient way of making our protein samples for Western Blots. She is convinced that heating at 50-70C for 10 minutes w/SDS is sufficient. I am convinced that samples need to be heated to at least 95C for 5 minutes w/SDS before proper denaturation can occur. The only reason I (personally) would use a slightly lower temperature is to avoid aggregation of multi-pass membrane proteins. Does anyone have any insight on this so we can settle our dispute? Thank you! kgambles Hi kgambles, Remember that your protein solution contains SDS and likely other denaturants, so there really is no need to heat the samples as high as 95oC. The combination of 70oC and denaturants for 10 minutes should be enough to denature a reasonably concentrated protein solution. Ivan Delgado ????o????????Y?υ???, ???sample protein ??????, loading buffer??SDS?????????. SDS is a detergent that is present in the SDS-PAGE sample buffer where, along with a bit of boiling, and a reducing agent (normally DTT or B-ME to break down protein-protein disulphide bonds), it disrupts the tertiary structure of proteins. This brings the folded proteins down to linear molecules. SDS also coats the protein with a uniform negative charge, which masks the intrinsic charges on the R-groups. SDS binds fairly uniformly to the linear proteins (around 1.4g SDS/ 1g protein), meaning that the charge of the protein is now approximately proportional to its molecular weight. |
10楼2017-06-02 18:50:24
3楼2017-05-26 13:41:54
果果gxl2013
金虫 (小有名气)
- 应助: 0 (幼儿园)
- 金币: 1379.8
- 散金: 35
- 红花: 1
- 帖子: 253
- 在线: 108.5小时
- 虫号: 3081153
- 注册: 2014-03-23
- 专业: 神经生物学
4楼2017-05-26 15:19:48
果果gxl2013
金虫 (小有名气)
- 应助: 0 (幼儿园)
- 金币: 1379.8
- 散金: 35
- 红花: 1
- 帖子: 253
- 在线: 108.5小时
- 虫号: 3081153
- 注册: 2014-03-23
- 专业: 神经生物学
5楼2017-05-26 15:20:38
6楼2017-05-26 18:48:34
果果gxl2013
金虫 (小有名气)
- 应助: 0 (幼儿园)
- 金币: 1379.8
- 散金: 35
- 红花: 1
- 帖子: 253
- 在线: 108.5小时
- 虫号: 3081153
- 注册: 2014-03-23
- 专业: 神经生物学
7楼2017-05-26 21:12:59
★ ★ ★ ★
小木虫: 金币+0.5, 给个红包，谢谢回帖
果果gxl2013: 金币+3 2017-06-02 15:08:07
小木虫: 金币+0.5, 给个红包，谢谢回帖
果果gxl2013: 金币+3 2017-06-02 15:08:07
|
我們小鼠肝組織, 也有用RIPA裂解, 然后12000 4度离心20分钟，取上清定量加LDS sample(loading) buffer+DTT(SDS sample buffer也可以)，70度10miin後冰上，-20度保存的。 https://www.scientistsolutions.c ... aking-wblot-samples |
8楼2017-05-31 14:09:20
果果gxl2013
金虫 (小有名气)
- 应助: 0 (幼儿园)
- 金币: 1379.8
- 散金: 35
- 红花: 1
- 帖子: 253
- 在线: 108.5小时
- 虫号: 3081153
- 注册: 2014-03-23
- 专业: 神经生物学
多有打扰，请多包涵9楼2017-06-02 15:11:44