Human TACC3 knockout HEK-293T cell line
- Advanced Validation
- What is this?
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(0 Publication)
- Sanger seq
Unknown
Sanger Sequencing - Human TACC3 knockout HEK-293T cell line (AB266316)
Homozygous : Insertion of the selection cassette in exon 4
- Cell Culture
Lab
Cell Culture - Human TACC3 knockout HEK-293T cell line (AB266316)
Representative images TACC3 knockout HEK293T cells, low and high confluency examples (top left and right respectively) and wild-type HEK293T 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 M5000 microscope.
- WB
Lab
Western blot - Human TACC3 knockout HEK-293T cell line (AB266316)
Blocking and diluting buffer and concentration : 5% NFDM/TBST
In Western blot, ab134154 was shown to bind specifically to TACC3. Target of interest was observed at 140 kDa in wild-type 293T cell lysates (lane 1) with no signal observed at this size in TACC3 knockout cell line (lane 2) (lane 2, knockout cell line ab266316).
The bands beneath the target band (140 kDa) are likely to be degraded target fragments.
ab181602 was used as a GAPDH loading control at 1/200000 dilution.
All lanes:
Western blot - Anti-TACC3 antibody [EPR7756] (<a href='/en-us/products/primary-antibodies/tacc3-antibody-epr7756-ab134154'>ab134154</a>) at 1/1000 dilution
Lane 1:
Wild-type 293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg
Lane 2:
Western blot - Human TACC3 knockout HEK-293T cell line (ab266316) at 20 µg
Lane 3:
HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution
Predicted band size: 90 kDa
Observed band size: 140 kDa,37 kDa
false
Exposure time: 81s
Complementary Products
Primary Antibodies
AB134154
RabMAb|Recombinant|KO Validated|20ul selling size
![Immunocytochemistry/ Immunofluorescence - Anti-TACC3 antibody [EPR7756] (AB134154)](https://content.abcam.com/products/images/tacc3-antibody-epr7756-ab134154--immunocytochemistry-immunofluorescence-img124253.jpg)
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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
TACC3 is part of a protein complex that contributes to mitosis. It collaborates with the Aurora A kinase and ch-TOG protein to ensure proper microtubule dynamics and spindle formation. This complex stabilizes kinetochore microtubules by connecting them to the centrosomes allowing accurate chromosome segregation during cell division. TACC3's actions are fundamental to cell cycle regulation and maintaining genomic stability.
Pathways
The role of TACC3 is linked to the cell cycle regulation and mitosis pathway. It interacts closely with the Aurora A kinase pathway playing a role in regulating centrosome maturation and spindle assembly. In these pathways proteins like TPX2 and PLK1 are also involved working alongside TACC3 to maintain the fidelity of cell division and ensure equal distribution of genetic material to daughter cells.
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
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