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: see datasheet
Kit contents
The SUMOylation 1 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-SUMOylation inhibitor Cocktail Precision Red™ Protein Assay Reagent | SUMOylation 1 Affinity Beads IP Control Beads
| BlastR™ Wash Buffer
| Spin Columns Bead Elution Buffer
| Chemiluminescent Reagent A Chemiluminescent Reagent B Anti-SUMO1 antibody
|
Example results
There are many applications for these kits, here we describe an interesting example:
Application 1: Investigate significant SUMOylation 1 events
Immunoprecipitating total SUMO1 profiles using the Signal-Seeker™ SUMOylation 1 Detection Kit compared to older SUMO1 tools (A) HAP1 wildtype (WT) or SUMO1 knockout (KO) lysate, was obtained using BlastR lysis and filter system. 1 mg of each lysate were incubated with 40 µg of each SUMO1 affinity reagent: ASM11-beads (Cytoskeleton), 21C7 (Invitrogen—purified), 21C7 (DSHB—supernatant), D11-beads (Santa Cruz) and conjugated SUMO1 IgG control beads (CIG03). 21C7 antibodies were captured with protein G agarose beads to enrich for SUMO-1 modified proteins. Samples were separated by SDS-PAGE and transferred to PVDF. Enriched SUMO1 samples were analyzed by western blot using ASM01 (Cytoskeleton) antibody at 1:5000, and mouse Trubelot Ultra-HRP secondary at 1:1000 in 5% milk. Trueblot secondary was used to minimize heavy and light chain detection from 21C7 samples. (B): IP was performed using ASM11 as in Fig 1A. SUMO1 modified proteins were visualized with ASM01 1:5000, and anti-mouse secondary at 1:20,000 to highlight the profile of SUMOylated proteins in the region between 64-30 kDa that may be masked by heavy and light chain interference when using unconjugated antibodies for IP. |
Immunoprecipitating SUMO1 modified target proteins using the Signal-Seeker™ SUMOylation 1 Detection Kit compared to older SUMO1 tools
HAP1 wildtype (WT) or SUMO1 knockout (KO) lysate, was obtained using BlastR lysis and filter system. 1 mg of each lysate were incubated with 40 µg of each SUMO1 affinity reagent: ASM11-beads (Cytoskeleton), 21C7 (DSHB—supernatant), and conjugated SUMO1 IgG control beads (CIG03). 21C7 antibodies were captured with protein G agarose beads to enrich for SUMO-1 modified proteins. Samples were separated by SDS-PAGE and transferred to PVDF. Target proteins: (A) TFII-I, RanGAP1, and (B) schmd1 were analyzed for their SUMO1 modified forms by western blot. Anti-rabbit-HRP labeled secondary antibody was used at 1:10,000. All three primary antibodies are rabbit polyclonal antibodies, and should not bind heavy and light chain fragments from the IP antibody.
• Pharmacological investigation of SUMOylating and de-SUMOylating enzymes involved in regulation of target proteins.
• Investigate SUMOylation under a variety of different growth factors or drug treatments.
• Examine the interaction of SUMOylated target proteins with its downstream effectors.
• Examine crosstalk between SUMOylation 1 and other PTMs for target proteins.
For more information contact: signalseeker@cytoskeleton.com
Associated Products:
Signal-Seeker™ SUMOylation 2/3 Detection Kit (Cat. # BK162)
Signal-Seeker™ Ubiquitination Detection Kit (Cat. # BK161)
Signal-Seeker™: BlastR™ Rapid Lysate Prep Kit (Cat. # BLR01)
Signal-Seeker™ SUMOylation 2/3 Affinity Beads (Cat.# ASM11-beads)
Signal-Seeker™: PTMtrue™ SUMOylation 2/3 Antibody (Cat.# ASM01)
Click on the pdf icon below to download the manual 
New product released in 2018! For the most recent publications citing this and other Signal-Seeker™ products, see our Signal-Seeker™ Validation Data Page click here
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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如果要是直接注射，就不知道了
两个意义不一样啊，定性是确定你抗体是阴还是阳的，如果阳证明2个情况，一个是你感染了丙肝病毒，另一个是你曾经感染过这个病毒，现在已经好了。但是到底是哪个情况还要进一步做定量测试既病毒RNA检测。如果这个测试值在最低线以下就证明你现在没事，不具有传染性也不是患者只是携带者；如果高于最低线那你就是患者了，就需要治疗
只要引物设计的可以在基因组上做，那么理论上是没问题的！
只需要用RT-PCR荧光定量试剂盒里面的第二个，专门用来做Real-timePCR的那些试剂就可以了！
现在跑tricine－sds－page电泳老是跑不好，
我刚做了bradford定量，
觉得差异与空菌对照还是挺明显的，
但是这个方法只能初略的估计，
还是不能精确定量，
做HPLC我现在还没有条件，
所以我想能不能用BCA或者lowry法先进行定量啊？
这种方法如果做出来结果的话，
能不能发个sci之类的文章啊？
希望高手们指点一下！
谢谢了
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至少可以说同一种蛋白定量试剂盒并不适用于所有细胞
蛋白定量试剂盒有分Bradford，BCA，Lowry等，每一种对不同的细胞都有独特的适用范围和灵敏度。所以需要多种蛋白定量试剂盒对比检测才能让结果准确
