Human DDIT3 knockout SW480 cell line
- Advanced Validation
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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 provided. Knockout achieved by CRISPR/Cas9 X = 1 bp deletion Frameshift: 98.84%.
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
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='/en-us/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='/en-us/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
- NGS
Supplier Data
Next Generation Sequencing - Human DDIT3 knockout SW480 cell line (AB269585)
1 bp deletion after Trp20 of the WT protein
- NGS
Supplier Data
Next Generation Sequencing - Human DDIT3 knockout SW480 cell line (AB269585)
Knockout achieved by CRISPR/Cas9; X = 1 bp deletion; Frameshift : 98.84%
Reactivity data
Product details
Recommended control: Human wild-type SW480 cell line (ab271146). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.
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.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
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 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.
Culture medium
Ham's F-12 + 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.
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
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-img444980.jpg)
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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