Human TRIB3 knockout HCT116 cell lysate
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TRIB3 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Homozygous: 35 bp deletion in exon 2.
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 HCT116 cell lysate (AB275249)
False colour image of Western blot : Anti-TRIB3 antibody 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, ab137526 was shown to bind specifically to TRIB3. A band was observed at 40 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in TRIB3 knockout cell line ab273718 (knockout cell lysate ab275249). To generate this image, wild-type and TRIB3 knockout HCT 116 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 °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-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes:
Western blot - Anti-TRIB3 antibody (<a href='/en-us/products/primary-antibodies/trib3-antibody-ab137526'>ab137526</a>) at 1/1000 dilution
Lane 1:
Wild-type HCT 116 cell lysate at 20 µg
Lane 2:
Western blot - Human TRIB3 knockout HCT116 cell lysate (ab275249) at 20 µg
Secondary
All lanes:
Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Predicted band size: 40 kDa
Observed band size: 40 kDa
false
- WB
Lab
Western blot - Human TRIB3 knockout HCT116 cell lysate (AB275249)
Lane 1 : Wild-type HCT116 cell lysate 20 ug
Lane 2 : TRIB3 knockout HCT116 cell lysate 20 ug
Lane 3 : Hap1 cell lysate 20 ug
Lane 4 : HeLa cell lysate 20 ug
Lanes 1 - 4 : Merged signal (red and green). Green - ab75846 observed at 40 kDa. Red - loading control, ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab75846 was shown to react with TRIB3 in wild-type HCT 116 cells in western blot with loss of signal observed in TRIB3 knockout cell line ab273718 (TRIB3 knockout cell lysate ab275249). HCT 116 wild-type and TRIB3 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab75846 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 10000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-TRIB3 antibody [EPR3151Y] (<a href='/en-us/products/primary-antibodies/trib3-antibody-epr3151y-ab75846'>ab75846</a>) at 1/10000 dilution
Lane 1:
Wild-type HCT116 cell lysate at 20 µg
Lane 2:
TRIB3 knockout HCT116 cell lysate at 20 µg
Lane 2:
Western blot - Human TRIB3 knockout HCT116 cell line (<a href='/en-us/products/cell-lines/human-trib3-knockout-hct116-cell-line-ab273718'>ab273718</a>)
Lane 3:
Hap1 cell lysate at 20 µg
Lane 4:
HeLa cell lysate at 20 µg
Predicted band size: 40 kDa
Observed band size: 40 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human TRIB3 knockout HCT116 cell lysate (AB275249)
Allele-1 : 35 bp deletion in exon 2 .
Reactivity data
Product details
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.
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
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: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
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'.
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