Human TRIP13 (PCH2) knockout HEK-293T cell line
- Advanced Validation
- What is this?
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TRIP13 KO cell line available to order. KO validated by. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 17 bp deletion in exon 1 and Insertion of the selection cassette in exon 1.
View Alternative Names
16E1-BP, HPV16 E1 protein-binding protein, Homo sapiens HPV16 E1 protein binding protein mRNA complete cds, Human papillomavirus type 16 E1 protein-binding protein, Pachytene checkpoint protein 2 homolog, TR-interacting protein 13, TRIP-13, TRP13_HUMAN, Thyroid hormone receptor interactor 13, Thyroid receptor-interacting protein 13
- WB
Lab
Western blot - Human TRIP13 (PCH2) knockout HEK-293T cell line (AB266495)
Anti-TRIP13 antibody (ab128171) staining at 1/2000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab128171 was shown to bind specifically to TRIP13. A band was observed at 48 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in TRIP13 knockout cell line. To generate this image, wild-type and TRIP13 knockout HEK-293T 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-TRIP13/PCH2 antibody (<a href='/en-us/products/primary-antibodies/trip13-pch2-antibody-ab128171'>ab128171</a>) at 1/2000 dilution
Lane 1:
Wild-type HEK-293T cell lysate at 20 µg
Lane 2:
Western blot - Human TRIP13 (PCH2) knockout HEK-293T cell line (ab266495)
Lane 2:
TRIP13 knockout HEK-293T cell lysate at 20 µg
Lane 2:
Western blot - Human TRIP13 (PCH2) knockout HEK-293T cell lysate (<a href='/en-us/products/cell-lysates/human-trip13-pch2-knockout-hek-293t-cell-lysate-ab258736'>ab258736</a>)
Lane 3:
HCT 116 cell lysate at 20 µg
Lane 4:
SW480 cell lysate at 20 µg
Predicted band size: 49 kDa
Observed band size: 48 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human TRIP13 (PCH2) knockout HEK-293T cell line (AB266495)
Allele-2 : Insertion of the selection cassette in exon 1.
- Sanger seq
Unknown
Sanger Sequencing - Human TRIP13 (PCH2) knockout HEK-293T cell line (AB266495)
Allele-1 : 17 bp deletion in exon 1
Reactivity data
Product details
Recommended control: Human wild-type HEK293T cell line (ab255449). 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, 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
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
TRIP13 contributes to efficient cell cycle progression by participating in the spindle assembly checkpoint (SAC). It forms part of the mitotic checkpoint complex ensuring that chromosomes are correctly attached to the spindle microtubules before progression through mitosis. TRIP13 works in conjunction with other proteins like MAD2 and BUB1 to maintain genomic stability.
Pathways
TRIP13 is involved in the cell cycle control and DNA repair pathways. It influences the mitotic spindle checkpoint interacting with proteins such as MAD2 which ensures that cells do not proceed to anaphase until all chromosomes reach proper alignment. TRIP13 also plays a role in homologous recombination repair by regulating the processing of recombination intermediates.
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
Primary Antibodies
AB313703
Anti-TRIP13/PCH2 antibody [EPR27063-80] - BSA and Azide free
RabMAb|KO Validated|Recombinant
![Immunoprecipitation - Anti-TRIP13/PCH2 antibody [EPR27063-80] - BSA and Azide free (AB313703)](https://content.abcam.com/products/images/trip13-pch2-antibody-epr27063-80-bsa-and-azide-free-ab313703--immunoprecipitation-img418671.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