人CDC42 knockout HEK-293T cell裂解物(ab256868)
概述
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产品名称
人CDC42 knockout HEK-293T cell裂解物
参阅全部 CDC42 试剂盒 -
产品概述
Knockout cell lysate achieved by CRISPR/Cas9. -
Parental Cell Line
HEK293T -
Organism
Human -
Mutation description
Knockout achieved by using CRISPR/Cas9, Homozygous: 2 bp insertion in exon 3. -
Passage number
<20 -
Knockout validation
Sanger Sequencing, Western Blot (WB) -
Reconstitution notes
To use as WB control, resuspend the lyophilizate in 50 µL of LDS* Sample Buffer to have a final concentration of 2 mg/ml. For reducing conditions, we recommend a final concentration of 0.1 M DTT.*Usage of SDS sample buffer is not recommended with these lyophilized lysates.
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说明
Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version - found here. Please refer to our lysis protocol for further details on how our lysates are prepared.
User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.
Access thousands of knockout cell lysates, generated from commonly used cancer cell lines.
See here for more information on knockout cell lysates.Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.
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经测试应用
适用于: WBmore details
性能
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存放说明
Store at -80°C. Please refer to protocols. -
组件 1 kit ab262823 - Human CDC42 knockout HEK293T cell lysate 1 x 100µg ab255553 - Human wild-type HEK293T cell lysate 1 x 100µg -
研究领域
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Cell type
epithelial -
STR Analysis
Amelogenin X D5S818: 8, 9 D13S317: 12, 14 D7S820: 11 D16S539: 9, 13 vWA: 16, 19 TH01: 7, 9.3 TPOX: 11 CSF1PO: 11, 12
靶标
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功能
Plasma membrane-associated small GTPase which cycles between an active GTP-bound and an inactive GDP-bound state. In active state binds to a variety of effector proteins to regulate cellular responses. Involved in epithelial cell polarization processes. Causes the formation of thin, actin-rich surface projections called filopodia. -
序列相似性
Belongs to the small GTPase superfamily. Rho family. CDC42 subfamily. -
翻译后修饰
AMPylation at Tyr-32 and Thr-35 are mediated by bacterial enzymes in case of infection by H.somnus and V.parahaemolyticus, respectively. AMPylation occurs in the effector region and leads to inactivation of the GTPase activity by preventing the interaction with downstream effectors, thereby inhibiting actin assembly in infected cells. It is unclear whether some human enzyme mediates AMPylation; FICD has such ability in vitro but additional experiments remain to be done to confirm results in vivo. -
细胞定位
Cell membrane. - Information by UniProt
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形式
There are 2 isoforms produced by alternative splicing. Isoform 1 also known as: Brain; Isoform 2 also known as: Placental. -
别名
- CDC42
- CDC42_HUMAN
- CDC42Hs
see all
相关产品
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KO cell lines
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab256868于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
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WB |
Use at an assay dependent concentration. Predicted molecular weight: 21 kDa.
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说明 |
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WB
Use at an assay dependent concentration. Predicted molecular weight: 21 kDa. |
图片
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Lane 1: Wild-type HEK-293T cell lysate (20 µg)
Lane 2: CDC42 knockout HEK-293T cell lysate (20 µg)
Lane 3: MCF7 cell lysate (20 µg)
Lane 4: HepG2 cell lysate (20 µg)
Lanes 1-4: Merged signal (red and green). Green - ab187643 observed at 20 kDa. Red - loading control ab8245 observed at 37 kDa.
ab187643 Anti-CDC42 antibody [EPR15620] was shown to specifically react with CDC42 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266522 (knockout cell lysate ab256868) was used. Wild-type and CDC42 knockout samples were subjected to SDS-PAGE. ab187643 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 5000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Lane 1: Wild-type HEK-293T cell lysate (20 µg)
Lane 2: CDC42 knockout HEK-293T cell lysate (20 µg)
Lane 3: MCF7 cell lysate (20 µg)
Lane 4: HepG2 cell lysate (20 µg)
Lanes 1-4: Merged signal (red and green). Green - ab64533 observed at 20 kDa. Red - loading control ab8245 observed at 37 kDa.
ab64533 Anti-CDC42 antibody was shown to specifically react with CDC42 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266522 (knockout cell lysate ab256868) was used. Wild-type and CDC42 knockout samples were subjected to SDS-PAGE. ab64533 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 µg/ml and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Homozygous: 2 bp insertion in exon 3
实验方案
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
数据表及文件
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SDS download
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Datasheet download
文献 (0)
ab256868 尚未被引用在任何文献中。