人E2F1 knockout HEK-293 cell裂解物(ab281354)
概述
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产品名称
人E2F1 knockout HEK-293 cell裂解物 -
产品概述
Knockout cell lysate achieved by CRISPR/Cas9.
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Parental Cell Line
HEK-293 -
Organism
Human -
Mutation description
Knockout achieved by CRISPR/Cas9; X = 17 bp deletion, 1 bp insertion; Frameshift = 99% -
Passage number
<20 -
Knockout validation
Next Generation Sequencing (NGS), 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 ab281394 - Human E2F1 knockout HEK-293 cell lysate 1 x 100µg ab259780 - Human wild-type HEK-293 cell lysate 1 x 100µg -
研究领域
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Cell type
epithelial -
Gender
Female
靶标
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功能
Transcription activator that binds DNA cooperatively with dp proteins through the E2 recognition site, 5'-TTTC[CG]CGC-3' found in the promoter region of a number of genes whose products are involved in cell cycle regulation or in DNA replication. The DRTF1/E2F complex functions in the control of cell-cycle progression from G1 to S phase. E2F-1 binds preferentially RB1 protein, in a cell-cycle dependent manner. It can mediate both cell proliferation and p53-dependent apoptosis. -
序列相似性
Belongs to the E2F/DP family. -
翻译后修饰
Phosphorylated by CDK2 and cyclin A-CDK2 in the S-phase.
Acetylation stimulates DNA-binding. Enhanced under stress conditions such as DNA damage and inhibited by retinoblastoma protein pRB. Regulated by KAP1/TRIM28 which recruits HDAC1 to E2F1 resulting in deacetylation. Acetylated by P/CAF/KAT2B. -
细胞定位
Nucleus. - Information by UniProt
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别名
- Dmel\CG6376
- Dmel_CG6376
- drosE2F1
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ab281354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 |
|---|---|---|
| WB |
Use at an assay dependent concentration.
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| 说明 |
|---|
|
WB
Use at an assay dependent concentration. |
图片
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Next Generation Sequencing - Human E2F1 knockout HEK-293 cell lysate (ab281354)Knockout achieved by CRISPR/Cas9; X = 17 bp deletion, 1 bp insertion; Frameshift = 99%
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Western blot - Human E2F1 knockout HEK-293 cell lysate (ab281354)Lane 1: Wild-type HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate 20 ug
Lane 2: E2F1 knockout HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate 20 ug
Lane 3: Hep G2 (Human liver hepatocellular carcinoma cell line) whole cell lysate 20 ug
Lane 4: U-2 OS cell lysate 20 ug
Lane 5: HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate 20 ug
Lanes 1 - 5: Merged signal (red and green). Green - ab4070 observed at 55 kDa. Red - loading control ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) observed at 37 kDa.
ab4070 was shown to react with E2F1 in wild-type HEK-293 cells in Western blot with loss of signal observed in E2F1 knockout cell line ab269502 (knockout cell lysate ab281354). Wild-type HEK-293 and E2F1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab4070 and ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) overnight at 4°C at a 1 in 500 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
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)
ab281354 尚未被引用在任何文献中。