AB269585

人DDIT3 knockout SW480 cell line

Human DDIT3 knockout SW480 cell line

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DDIT3 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control available. Knockout achieved by CRISPR/Cas9 X = 1 bp deletion Frameshift: 98.84%. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.

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

3 Images

Supplier Data

Western blot - Human DDIT3 knockout SW480 cell line (AB269585)

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 HeLa Vehicle Control Tunicamycin (20ug/mL, 4h) cell lysate at 20 µg

Lane 2:

Wild-type HeLa Treated Tunicamycin (20ug/mL, 4h) cell lysate at 20 µg

Lane 3:

DDIT3 knockout HeLa Vehicle Control Tunicamycin (20ug/mL, 4h) cell lysate at 20 µg

Lanes 3 - 4:

Western blot - Human DDIT3 knockout SW480 cell line (ab269585)

Lanes 3 - 4:

Western blot - Human DDIT3 knockout HeLa cell lysate (<a href='/products/cell-lysates/human-ddit3-knockout-hela-cell-lysate-ab256889'>ab256889</a>)

Lane 4:

DDIT3 knockout HeLa Treated Tunicamycin (20ug/mL, 4h) cell lysate at 20 µg

Predicted band size: 19 kDa

Observed band size: 25 kDa

false

Supplier Data

Next Generation Sequencing - Human DDIT3 knockout SW480 cell line (AB269585)

1 bp deletion after Trp20 of the WT protein

Supplier Data

Next Generation Sequencing - Human DDIT3 knockout SW480 cell line (AB269585)

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

形式

Liquid

form

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": { "NGS": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" }, "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }

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产品详情

We will provide viable cells that proliferate on revival.

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

Dry Ice

处理步骤

初始处理指南

Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.

1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO₂. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x10⁴ cells/cm². Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.

传代培养指南

All seeding densities should be based on cell counts gained by established methods.
A guide seeding density of 2-3x10⁴ cells/cm² is recommended.
Cells should be passaged when they have achieved 80-90% confluence.
Slow growing. A partial media change is recommended at least twice between passages.

培养基

Ham's F-12 + 10% FBS

低温储藏试剂

Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

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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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靶点信息

See full target information DDIT3

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Abcam Product Promise

我们致力于为您的研究提供高质量的试剂，为您科研的每一步提供支持。若我们的产品未能达到预期性能，我们向您提供 Abcam Product Promise 保障。

详情请参阅我们的条款与条件。

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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人DDIT3 knockout SW480 cell line

Human DDIT3 knockout SW480 cell line

Western blot - Human DDIT3 knockout SW480 cell line (AB269585)

Next Generation Sequencing - Human DDIT3 knockout SW480 cell line (AB269585)

Next Generation Sequencing - Human DDIT3 knockout SW480 cell line (AB269585)