AB270708

人DDIT3 knockout SW480 cell裂解物

Name: 人DDIT3 knockout SW480 cell lysate (ab270708)| Abcam中文官网
Brand: Abcam Limited
SKU: AB270708
Price: 6405 CNY
Availability: InStock

Human DDIT3 knockout SW480 cell lysate

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DDIT3 KO cell lysate available now. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control included. Knockout achieved by CRISPR/Cas9 X = 1 bp deletion Frameshift: 98.84%.

查看别名

C/EBP Homology Protein, C/EBP zeta, C/EBP-homologous protein, C/EBP-homologous protein 10, CCAAT/enhancer binding protein homologous protein, CHOP, CHOP-10, DDIT3_HUMAN, DNA Damage Inducible Transcript 3, DNA damage-inducible transcript 3 protein, GADD 153, Growth Arrest and DNA Damage Inducible Protein 153, Growth arrest and DNA damage-inducible protein GADD153, MGC4154

2 Images

Lab

Western blot - Human DDIT3 knockout SW480 cell lysate (AB270708)

Lane 1 : Wild-type SW480 cell lysate 20 ug

Lane 2 : DDIT3 knockout SW480 cell lysate 20 ug

Lane 3 : Untreated HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate 20 ug

Lane 4 : HeLa + DMSO control cell lysate 20 ug

Lane 5 : HeLa + tunicamycin (20ug/mL,4 hours) cell lysate 20 ug

Lanes 1 - 5 : Merged signal (red and green). Green - ab11419 observed at 26 kDa. Red - loading control ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) observed at 55kDa.

ab11419 was shown to react with DDIT3 in wild-type SW480 cells in western blot with loss of signal observed in DDIT3 knockout cell line ab269585 (knockout cell lysate ab270708). Wild-type and DDIT3 knockout SW480 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween^®) before incubation with ab11419 and ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) overnight at 4°C at 5 μg/ml 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 hour at room temperature before imaging.

All lanes:

Western blot - Anti-CHOP antibody [9C8] (<a href='/products/primary-antibodies/chop-antibody-9c8-ab11419'>ab11419</a>) at 5 µg/mL

Lane 1:

Wild-type SW480 cell lysate at 20 µg

Lane 2:

DDIT3 knockout SW480 cell lysate at 20 µg

Lane 3:

Untreated HeLa cell lysate at 20 µg

Lane 4:

HeLa + DMSO control cell lysate at 20 µg

Lane 5:

HeLa + tunicamycin (20ug/mL,4 hours) cell lysate at 20 µg

Predicted band size: 19 kDa

Observed band size: 26 kDa

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Supplier Data

Next Generation Sequencing - Human DDIT3 knockout SW480 cell lysate (AB270708)

Knockout achieved by CRISPR/Cas9; X = 1 bp deletion; Frameshift : 98.84%

关键信息

细胞类型

SW480

种属

Human

组织

Colon

敲除验证

Next Generation Sequencing,Western blot

突变描述

Knockout achieved by CRISPR/Cas9 X = 1 bp deletion Frameshift: 98.84%

疾病

Adenocarcinoma

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反应性数据

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }

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产品详情

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
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.

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. 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.

This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.

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规格

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性能和储存信息

基因名称

DDIT3

基因编辑类型

Knockout

基因编辑方法

CRISPR technology

敲除验证

Next Generation Sequencing, Western blot

运输条件

Ambient - Can Ship with Ice

补充信息

This supplementary information is collated from multiple sources and compiled automatically.

DDIT3 also known as CHOP (C/EBP homologous protein) is a transcription factor commonly expressed in cells under stress conditions such as endoplasmic reticulum stress. It has a molecular weight of approximately 29 kDa. As part of the system that manages cell stress responses DDIT3 expression can occur in various tissues but it is particularly notable in the context of cellular stress regulation. Researchers often use DDIT3 immunohistochemistry to study its presence and expression pattern in different tissues.

Biological function summary

DDIT3 plays a significant role in mediating cellular stress responses and promoting apoptosis when adaptive pathways fail. DDIT3 is not typically part of a multi-subunit complex but it acts in concert with other stress-related proteins to modulate gene expression. By inducing apoptosis DDIT3 helps remove severely damaged cells maintaining overall tissue health. However excessive DDIT3 activity under prolonged stress can lead to cell loss and tissue damage.

Pathways

DDIT3 is involved in the unfolded protein response (UPR) and endoplasmic reticulum stress pathways. It interacts with proteins such as ATF4 and PERK which are part of the mechanism that governs these pathways. DDIT3 induction contributes to the UPR pathway balancing the cell's fate between repair and apoptosis. This balance is important for cell survival under stress and avoids detrimental cellular damage.

DDIT3 has a strong connection with diabetes and neurodegenerative diseases. In diabetes DDIT3 overactivation can result in pancreatic beta-cell apoptosis worsening the disease's progression. Furthermore its involvement in neurodegenerative diseases includes its association with the accumulation of misfolded proteins exacerbating cellular stress conditions. DDIT3's interaction with proteins such as BCL2 further influences disease pathways by enhancing apoptotic signals.

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细胞培养

生物安全等级

EU: 1 US: 1

贴壁/悬浮

Adherent

性别

Male

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产品实验方案

Visit the General protocols
Visit the Troubleshooting

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人DDIT3 knockout SW480 cell裂解物

Human DDIT3 knockout SW480 cell lysate

Western blot - Human DDIT3 knockout SW480 cell lysate (AB270708)

Next Generation Sequencing - Human DDIT3 knockout SW480 cell lysate (AB270708)