Human DDIT3 knockout HeLa cell line
- Advanced Validation
- What is this?
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DDIT3 KO cell line available to order. KO validated by. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 2. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.
View Alternative Names
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
- WB
Lab
Western blot - Human DDIT3 knockout HeLa cell line (AB265760)
False colour image of Western blot : Anti-DDIT3 antibody [9C8] staining at 5 µg/ml, shown in green; Rabbit anti-alpha Tubulin antibody [EP1332Y] (ab52866) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab11419 was shown to bind specifically to DDIT3. A band was observed at 25 kDa in wild-type y cell lysates with no signal observed at this size in DDIT3 knockout cell line ab265760 (knockout cell lysate ab256889). To generate this image, wild-type and DDIT3 knockout y cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged.Secondary antibodies used were Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) at 1/20000 dilution.
All lanes:
Western blot - Anti-CHOP antibody [9C8] (<a href='/en-us/products/primary-antibodies/chop-antibody-9c8-ab11419'>ab11419</a>) at 5 µg/mL
Lane 1:
Wild-type HeLa Vehicle Control Tunicamycin cell lysate at 20 µg
Lane 2:
Wild-type HeLa Treated Tunicamycin cell lysate at 20 µg
Lane 3:
DDIT3 knockout HeLa Vehicle Control Tunicamycin cell lysate at 20 µg
Lanes 3 - 4:
Western blot - Human DDIT3 knockout HeLa cell line (ab265760)
Lanes 3 - 4:
Western blot - Human DDIT3 knockout HeLa cell lysate (<a href='/en-us/products/cell-lysates/human-ddit3-knockout-hela-cell-lysate-ab256889'>ab256889</a>)
Lane 4:
DDIT3 knockout HeLa Treated Tunicamycin cell lysate at 20 µg
Predicted band size: 19 kDa
Observed band size: 25 kDa
false
- Cell Culture
Unknown
Cell Culture - Human DDIT3 knockout HeLa cell line (AB265760)
Representative images of DDIT3 knockout HeLa cells, low and high confluency examples (top left and right respectively) and wild-type HeLa cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS XL Core microscope.
- Sanger seq
Unknown
Sanger Sequencing - Human DDIT3 knockout HeLa cell line (AB265760)
Homozygous : 1 bp insertion in exon 2.
Reactivity data
Product details
We will provide viable cells that proliferate on revival.
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.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Zygosity
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Handling procedures
Initial handling guidelines
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.
Subculture guidelines
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 2x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
Culture medium
DMEM (High Glucose) + 10% FBS
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
Complementary Products
![Immunocytochemistry/ Immunofluorescence - Anti-CHOP antibody [9C8] (AB11419)](https://content.abcam.com/products/images/chop-antibody-9c8-ab11419--immunocytochemistry-immunofluorescence-img77075.jpg)
Primary Antibodies
AB229912
Anti-PERK antibody [EPR19876-294]
20ul selling size|RabMAb|Recombinant|KO Validated
![Western blot - Anti-PERK antibody [EPR19876-294] (AB229912)](https://content.abcam.com/products/images/perk-antibody-epr19876-294-ab229912--western-blot-img14494.jpg)
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com