Human PTTG1 (Securin) knockout HEK-293T cell line
- Advanced Validation
- What is this?
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(0 Publication)
- WB
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Western blot - Human PTTG1 (Securin) knockout HEK-293T cell line (AB266231)
Lanes 1-3 : Merged signal (red and green). Green - ab26273 observed at 28 kDa. Red - loading control ab8245 observed at 37 kDa.
ab26273 Anti-Securin antibody was shown to specifically react with Securin in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266231 (knockout cell lysate ab257289) was used. Wild-type and Securin knockout samples were subjected to SDS-PAGE. ab26273 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-Securin antibody (<a href='/en-us/products/primary-antibodies/securin-antibody-ab26273'>ab26273</a>) at 1/500 dilution
Lane 1:
Wild-type HEK293T cell lysate at 20 µg
Lane 2:
PTTG1 knockout HEK293T cell lysate at 20 µg
Lane 2:
Western blot - Human PTTG1 (Securin) knockout HEK-293T cell line (ab266231)
Lane 3:
Daudi cell lysate at 20 µg
Secondary
All lanes:
Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/10000 dilution
Predicted band size: 22 kDa
Observed band size: 28 kDa
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- Cell Culture
Unknown
Cell Culture - Human PTTG1 (Securin) knockout HEK-293T cell line (AB266231)
Representative images of PTTG1 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 XL Core microscope.
- Sanger seq
Unknown
Sanger Sequencing - Human PTTG1 (Securin) knockout HEK-293T cell line (AB266231)
Homozygous : 8 bp deletion in exon 1
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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 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
Quality control - STR analysis
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
Securing proper chromosome segregation Securin forms a complex with Separase. By inhibiting Separase Securin ensures that separation of chromatids occurs only at the correct stage of mitosis. This regulatory function is critical for maintaining genomic stability and avoiding aneuploidy which can lead to tumorigenesis. The presence of Securin in dividing cells highlights its role in the cell cycle.
Pathways
Securin functions are integrated within the cell cycle and apoptosis pathways. In the cell cycle pathway Securin controls the metaphase-anaphase transition. This involves interaction with cyclin-dependent kinases (CDKs) and the anaphase-promoting complex (APC/C). In terms of apoptosis Securin may interact with proteins like Bcl-2 impacting cell survival and death decisions under stress conditions.
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
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. <p>1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.<br>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.<br>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.<br>4. Incubate the culture at 37°C incubator with 5% CO<sub>2</sub>. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.<br>5. Once confluent passage into an appropriate flask at a density of 2x10<sup>4</sup> cells/cm<sup>2</sup>. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.</p>
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.
Target data
Alternative Names
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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