人PIK3R1 (PI 3 Kinase p85 alpha) knockout HeLa cell裂解物(ab257029)
概述
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产品名称
人PIK3R1 (PI 3 Kinase p85 alpha) knockout HeLa cell裂解物
参阅全部 PI 3 Kinase p85 alpha 试剂盒 -
产品概述
Knockout cell lysate achieved by CRISPR/Cas9. -
Parental Cell Line
HeLa -
Organism
Human -
Mutation description
Knockout achieved by using CRISPR/Cas9, Insertion of the selection cassette in exon10. -
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 ab261938 - Human PIK3R1 knockout HeLa cell lysate 1 x 100µg ab255929 - Human wild-type HeLa cell lysate 1 x 100µg -
研究领域
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Cell type
epithelial -
Disease
Adenocarcinoma -
Gender
Female -
STR Analysis
Amelogenin X D5S818: 11, 12 D13S317: 12, 13.3 D7S820: 8, 12 D16S539: 9, 10 vWA: 16, 18 TH01: 7 TPOX: 8,12 CSF1PO: 9, 10
靶标
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功能
Binds to activated (phosphorylated) protein-Tyr kinases, through its SH2 domain, and acts as an adapter, mediating the association of the p110 catalytic unit to the plasma membrane. Necessary for the insulin-stimulated increase in glucose uptake and glycogen synthesis in insulin-sensitive tissues. -
组织特异性
Isoform 2 is expressed in skeletal muscle and brain, and at lower levels in kidney and cardiac muscle. Isoform 2 and isoform 4 are present in skeletal muscle (at protein level). -
序列相似性
Belongs to the PI3K p85 subunit family.
Contains 1 Rho-GAP domain.
Contains 2 SH2 domains.
Contains 1 SH3 domain. -
结构域
The SH3 domain mediates the binding to CBLB, and to HIV-1 Nef. -
翻译后修饰
Polyubiquitinated in T-cells by CBLB; which does not promote proteasomal degradation but impairs association with CD28 and CD3Z upon T-cell activation.
Phosphorylated. Dephosphorylated by PTPRJ. - Information by UniProt
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别名
- GRB1
- p85
- p85 alpha
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ab257029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: 83 kDa.
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说明 |
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WB
Use at an assay dependent concentration. Predicted molecular weight: 83 kDa. |
图片
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Lane 1: Wild-type HeLa cell lysate (20µg)
Lane 2: PIK3R1 knockout HeLa cell lysate (20µg)
Lanes 1- 2: Merged signal (red and green). Green - ab133595 observed at 90 kDa. Red - loading control ab8245 observed at 37 kDa.
ab133595 Recombinant Anti-PI 3 Kinase p85 alpha antibody [EPR5513] was shown to specifically react with PI 3 Kinase p85 alpha in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265116 (knockout cell lysate ab257029) was used. Wild-type and PI 3 Kinase p85 alpha knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab133595 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 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. -
Lane 1: Wild-type HeLa cell lysate (20µg)
Lane 2: PIK3R1 knockout HeLa cell lysate (20µg)
Lanes 1- 2: Merged signal (red and green). Green - ab191606 observed at 90 kDa. Red - loading control ab8245 observed at 37 kDa.
ab191606 Anti-PI 3 Kinase p85 alpha antibody [EPR18702] was shown to specifically react with PI 3 Kinase p85 alpha in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265116 (knockout cell lysate ab257029) was used. Wild-type and PI 3 Kinase p85 alpha knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab191606 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 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. -
Allele-1: Insertion of the selection cassette in exon10
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Allele-2: Insertion of the selection cassette in exon10
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)
ab257029 尚未被引用在任何文献中。