The Signal-Seeker™ line of produts have been developed to allow simple analysis of key regulatory protein modifications by specialists and non-specialists alike. The comprehensive Signal-Seeker™ kits provide an affinity bead system to isolate and enrich modified proteins from any given cell or tissue lysate. The enriched protein population is then analyzed by standard western blot procedures using a primary antibody to the target protein
Product Uses Include
- Investigate transient regulatory mechanisms
- Measure signalling events of multiple pathway member proteins
- Discover new modifications of your protein of interest
- Gain insight into regulatory mechanisms
- Measure endogenous or transiently expressed protein signalling events
Validation Data: Ubiquitination Detection Kit White Paper
Kit contents
The Ubiquitination kit contains the following components:
Lysis and protein quantitation step | IP and pre-clear step | Wash step | Elution step | Western step |
BlastR™ Lysis Buffer BlastR™ Dilution Buffer BlastR™ Filters Protease Inhibitor Cocktail De-Ubiquitinase inhibitor Cocktail Precision Red™ Protein Assay Reagent | Ubiquitination Affinity Beads Ubiquitination Control Beads
| BlastR™ Wash Buffer
| Spin Columns Bead Elution Buffer
| Chemiluminescent Reagent A Chemiluminescent Reagent B Anti-ubiquitin-HRP antibody
|
Example results
There are many applications for these kits (see next section below), here we describe an interesting application for Rac1 protein:
Application 1: Investigate highly transient regulation events
Figure 1: Swiss 3T3 cells were pre-treated with MG-132, then either untreated or treated with CNF1 (CN04) for 3 hours prior to lysis with BlastR buffer. The BK161 kit was utilized to perform the IP on 300 μg of lysate per condition. Lane 1: 3 μg input untreated lysate, Lane 2: Ubiquitin Affinity beads (UBA01) plus untreated lysate, Lane 3: UBA01 plus CN04 treated lysate, Lane 4: ubiquitin control beads (CUB02) plus untreated lysate, Lane 5: CUB02 plus CN04 treated lysate. Samples were analyzed for Rac1 ubiquitination using an anti-Rac1 antibody.
For more information contact: signalseeker@cytoskeleton.com
Associated Products:
Signal-Seeker™ Phosphotyrosine Detection Kit (Cat. # BK160)
Signal-Seeker™ SUMOylation 2/3 Detection Kit (Cat. # BK162)
Signal-Seeker™: BlastR™ Rapid Lysate Prep Kit (Cat. # BLR01)
Signal-Seeker™ Ubiquitin Affinity Beads (Cat.# UBA01-beads)
Signal-Seeker™: PTMtrue™ Ubiquitin Antibody (Cat.# AUB01)
Click on the pdf icon below to download the manual
Visit our Signal-Seeker™ Tech Tips and FAQs page for technical tips and frequently asked questions regarding this and other Signal-Seeker™ products click here
If you have any questions concerning this product, please contact our Technical Service department at tservice@cytoskeleton.com
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蛋白的类型为PDGF。其有一种二聚体为PDGF-BB。
请教第一点:我看了Elisa的说明书,现在检测PDGF有两种试剂盒,一种是检测PDGF的,要求的试剂样本为组织匀浆;一种是检测PDGF-BB的,要求的待测样本为血液或血浆。
我现在的实验目的是想更精确,检验组织中的PDGF-BB,但是手中的是人体韧带组织样本,非PDGF-BB试剂盒所要求的组织样本为血浆或血清,我不知道是否可以用PDGF-BB的ELISA试剂盒的来做,所以特地请教:)
请教第二点:市面上有RB公司的96T,2500元;另外有Uscnlife的Elisa试剂盒,96T的3680元,都说是原装进口的,我该如何选择呢?为何差价这么多?谢谢指点!
一、免疫组化试剂盒一般包括:
1、特异性的一抗
2、免疫组化检测系统。
3、有的同时还具备显色系统。
二、不同的公司内容有差别,所以购买时一定了解清楚:
1、一抗是做人的还是兔、鼠的组织;
2、检测系统是ABC/SP/非生物素/其他;
3、试剂盒的内容都有什么?有没有显色系统?
4、如果没有,自己根据前面的检测系统需配哪个显色系统。(其他的辅助试剂如PBS缓冲液、抗原修复液等一般另卖或自配)
三、免疫组化抗体即指特异性一抗,是用来标记的指标,免疫组化过程的其他试剂一律另外配备。
另外:做实验时,如遇出结果困难,记得做阳性片对照,敢于怀疑检测系统或显色系统的问题。
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