Host/IsotypeMouse / IgG2b
Clone5D8B16
Species ReactivityBroad Reactivity
Validation Datasee datasheet
Anti-SUMO1 antibody, ASM11, is a SUMO1 mouse monoclonal antibody that is part of the Signal-Seeker™ product line. Anti-SUMO1 mouse monoclonal antibody was raised against full-length recombinant SUMO1 protein (Uniprot: P63165). The antibody has been shown to recognize a broad profile of SUMO1 proteins in HAP1, HeLa, and 293 cell lysates (Fig. 1A) and to detect sub-nanogram amounts of recombinant SUMO1 (Fig. 1B). The antibody shows high specificity (Fig. 1A WT vs KO lane).with no cross reactivity to SUMO2 (Fig. 1B). Furthermore, the sensitivity of ASM01 was compared to commercially available SUMO1 antibodies, and the data shows that ASM01 detects a more robust and specific profile of SUMO1 modified proteins (Fig. 1C). ASM01 is purified by Protein G affinity chromatography and is supplied as a lyophilized white powder. Each Lot of antibody is quality controlled to provide a high batch to batch consistency. The Lot specific µg per tube can be found in the Lot specific COA documents. |
Figure 1: Western Blot using SUMO-2/3 Antibody ASM01 was used at a 1:5000 dilution (0.2 µg/mL) following the recommended western blot protocol (see above). Figure 1A: HAP1 wildtype (WT) or SUMO1 knockout (KO) lysate, HeLa cell +/- NEM lysate and 293 cell +/- NEM lysate was obtained using BlastR lysis and filter system. 10 µg of each lysate were separated by SDS-PAGE and transferred to PVDF. Robust SUMO1 profiles were detected for every cell type. Specificity is shown by the lack of SUMO1 detection in SUMO1 KO cells and significantly diminished profiles in lysates prepared in the absence of the SENP inhibitor (NEM). Figure 1B: Titration of recombinant SUMO1 lanes 1-6 contain 5.0, 2.5, 1.25, 0.6, 0.3, and 0.15 ng SUMO1, while lane 7 contains 1000 ng of recombinant SUMO2. Figure 1C: HAP1 WT and KO lysate as prepared in 1A was used. 10 µg of each lysate were separated by SDS-PAGE and transferred to PVDF. SUMO1 proteins were detected using the recommended concentrations of ASM01 (Cytoskeleton), 21C7 (Invitrogen—purified), 21C7 (DSHB—supernatant), and D11 (Santa Cruz) as shown in the figure. All samples were developed and imaged simultaneously to ensure identical experimental conditions. To see the full Western blot protocol, see the product datasheet. |
Figure 2: Immunoprecipitation using SUMO-2/3 Antibody
HAP1 wildtype (WT) and knockout (KO) cells were prepared in BlastR buffer and filter system supplemented with NEM and PIC02. Figure 2A: 1 mg of lysate per reaction was used to immunoprecipitate (IP) SUMO1 modified proteins using 40 µg of 5D8 antibody (ASM01), 40 µg of ASM11-beads, 40 µg of 21C7 (DSHB-supernatant), or 40 µg of mouse IgG control antibody. 5D8, 21C7 supernatant, and mouse IgG were bound to Protein G Agarose as described in the method (see above). Enriched SUMO1 samples were analyzed by western blot using ASM01 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.
Figure 2B: IP was performed using ASM11 as in Fig 2A. 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. To see the full Immunoprecipitation protocol, see the product datasheet.
Immunofluorescence applications for ASM01 have not been tested
For more information contact: signalseeker@cytoskeleton.com
Associated Products:
Signal-Seeker™ SUMOylation 1 Detection Kit (Cat. # BK165)
Signal-Seeker™ SUMOylation 1 Affinity Beads (Cat.# ASM11-beads)
Signal-Seeker™: BlastR™ Rapid Lysate Prep Kit (Cat. # BLR01)
For product Datasheets and MSDSs please click on the PDF links below.
Sample Size Datasheet (Cat. ASM01-S):
Certificate of Analysis: available upon request
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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以下来自摆渡百科
细胞在发生凋亡时,会激活一些DNA内切酶,这些内切酶会切断核小体间的基因组DNA。细胞凋亡时抽提DNA进行电泳检测,可以发现180-200bp的DNA ladder。基因组DNA断裂时,暴露的3’-OH可以在末端脱氧核苷酸转移酶(Terminal
Deoxynucleotidyl Transferase, TdT) 的催化下加上荧光素 (FITC) 标记的dUTP
(fluorescein-dUTP) ,从而可以通过荧光显微镜或流式细胞仪进行检测,这就是TUNEL (TdT-mediated dUTP
Nick-End Labeling) 法检测细胞凋亡的原理。
第二个问题:兔子不能免疫兔子的。免疫的一个重要概念是识别“自己”和“非己”,如果对自己的蛋白产生免疫反应,那就麻烦了。
可以类比器官移植,亲缘关系越近,越不容易产生免疫排斥。
sc-153是兔抗大鼠ERK2的多克隆抗体
还有很多关于ERK1和ERK2的抗体
若想知道更多信息,你可以拨打Santa cruz 上海分公司的电话咨询
021-6093-6351
我们会按照你的实验需求推荐最适合你的产品。
想买消炎药怎么办?
去药店买药想买不是抗生素的消炎药,那工作人员说不是抗生素怎么消炎呢,请问知情网友求一替代品,谢了。
你看这个抗体的质量怎么样,说明里面有没有说可以做组化,还是只能做western blot。
暂无品牌问答