Human TRIB3 knockout HeLa cell line
- Advanced Validation
- What is this?
Be the first to review this product! Submit a review
|
(0 Publication)
TRIB3 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 2 bp deletion 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
C20orf97, NIPK, Neuronal cell death-inducible putative kinase, SINK, SKIP 3, TRB-3, TRIB3_HUMAN, Tribbles homolog 3, Tribbles pseudokinase 3, Tribbles3, p65-interacting inhibitor of NF-kappa-B
- WB
Lab
Western blot - Human TRIB3 knockout HeLa cell line (AB264792)
False colour image of Western blot : Anti-TRIB3 antibody [EPR3150] staining at 1/1000 dilution shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution shown in red. In Western blot ab75774 was shown to bind specifically to TRIB3. A band was observed at 40 kDa in wild-type HeLa cell lysates with no signal observed at this size in TRIB3 knockout cell line ab264792 (knockout cell lysate ab257765). To generate this image wild-type and TRIB3 knockout HeLa cell lysates were analysed. First samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°. 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-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
All lanes:
Western blot - Anti-TRIB3 antibody [EPR3150] (<a href='/en-us/products/primary-antibodies/trib3-antibody-epr3150-ab75774'>ab75774</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
TRIB3 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human TRIB3 knockout HeLa cell line (ab264792)
Predicted band size: 40 kDa
Observed band size: 40 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human TRIB3 knockout HeLa cell line (AB264792)
Homozygous : 2 bp deletion 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 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
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
The modulation of metabolic pathways and stress responses involves TRIB3 acting in several mechanisms. While TRIB3 itself is not part of a complex it exerts its effects by binding to and inhibiting the activity of other proteins notably AKT1 and several transcription factors. This modulation influences glucose metabolism and cellular response to unfolded protein stress indicating TRIB3's role as a significant regulator of cellular homeostasis.
Pathways
The influence of TRIB3 on insulin signaling and the unfolded protein response is significant. Within the insulin signaling pathway TRIB3 interacts with AKT1 a central protein that regulates glucose uptake and cell survival. This interaction inhibits AKT1's activity impacting glucose metabolism. In the unfolded protein response pathway TRIB3 impacts stress response regulation through interactions that modulate transcriptional activity further highlighting its regulatory roles.
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
![Immunoprecipitation - Anti-TRIB3 antibody [EPR3151Y] (AB75846)](https://content.abcam.com/products/images/trib3-antibody-epr3151y-ab75846--immunoprecipitation-img178490.jpg)
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