人GPX4 knockout HeLa cell裂解物(ab263935)
概述
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产品名称
人GPX4 knockout HeLa cell裂解物
参阅全部 Glutathione Peroxidase 4 试剂盒 -
产品概述
Knockout cell lysate achieved by CRISPR/Cas9.
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Parental Cell Line
HeLa -
Organism
Human -
Mutation description
Knockout achieved by CRISPR/Cas9; X = 26 bp deletion, 2 bp insertion; Frameshift: 93.31% -
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 ab280488 - Human GPX4 knockout HeLa cell lysate 1 x 100µg ab269597 - Human wild-type HeLa cell lysate 1 x 100µg -
研究领域
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Cell type
epithelial -
Disease
Adenocarcinoma -
Gender
Female
靶标
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功能
Protects cells against membrane lipid peroxidation and cell death. Required for normal sperm development and male fertility. Could play a major role in protecting mammals from the toxicity of ingested lipid hydroperoxides. Essential for embryonic development. Protects from radiation and oxidative damage. -
组织特异性
Present primarily in testis. -
序列相似性
Belongs to the glutathione peroxidase family. -
细胞定位
Mitochondrion. Cytoplasm. - Information by UniProt
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别名
- Glutathione peroxidase 4
- GPX 4
- GPX-4
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ab263935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: 22 kDa.
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说明 |
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WB
Use at an assay dependent concentration. Predicted molecular weight: 22 kDa. |
图片
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Knockout achieved by CRISPR/Cas9; X = 26 bp deletion, 2 bp insertion; Frameshift: 93.31%
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Lanes 1: Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate, 20 ug
Lanes 2: GPX4 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate, 20 ug
Lanes 3: Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate, 20 ug
Lanes 3: Hep G2 (Human liver hepatocellular carcinoma cell line) whole cell lysate, 20 ugab206266 was shown to react with Glutathione Peroxidase 4 (HRP) in wild-type HeLa cells in western blot. Loss of signal was observed when GPX4 knockout cell line ab262509 (knockout cell lysate ab263935) was used. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab206266 overnight at 4 °C at a 1 in 5000 dilution Blots were developed with Optiblot ECL reagent (ab133456) and imaged.
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Lanes 1: Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate, 20 ug
Lanes 2: GPX4 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate, 20 ug
Lanes 3: Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate, 20 ugLanes 1 - 3: Merged signal (red and green). Green - ab125066 observed at 20 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.
ab125066 was shown to react with Glutathione Peroxidase 4 in wild-type HeLa cells in Western blot with loss of signal observed in GPX4 knockout cell line ab262509 (knockout cell lysate ab263935). Wild-type HeLa and GPX4 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab125066 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
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Lanes 1: Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate, 20 ug
Lanes 2: GPX4 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate, 20 ug
Lanes 3: Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate, 20 ugLanes 1 - 3: Merged signal (red and green). Green - ab41787 observed at 20 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.
ab41787 was shown to react with Glutathione Peroxidase 4 in wild-type HeLa cells in Western blot with loss of signal observed in GPX4 knockout cell line ab262509 (knockout cell lysate ab263935). Wild-type HeLa and GPX4 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab41787 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at 1 µg/ml and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) 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)
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