Human SFN (14-3-3 sigma) knockout HeLa cell line
- Advanced Validation
- What is this?
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SFN KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 1 and 2 bp deletion in exon 1.
View Alternative Names
14-3-3 protein sigma, 1433S_HUMAN, ER, Epithelial cell marker protein 1, MGC143283, Mme1, OTTHUMP00000004242, RP23 137L22.11, SFN, SFN protein, Stratifin, YWHAS
- WB
Unknown
Western blot - Human SFN (14-3-3 sigma) knockout HeLa cell line (AB261779)
Lanes 1-3 : Merged signal (red and green). Green - ab14123 observed at 28 kDa. Red - loading control, ab52866 observed at 50 kDa.
ab14123 Anti-14-3-3 sigma/SFN antibody [1433S01] was shown to specifically react with 14-3-3 sigma/SFN in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261779 (knockout cell lysate ab257288) was used. Wild-type and 14-3-3 sigma/SFN knockout samples were subjected to SDS-PAGE. ab14123 and Anti-alpha Tubulin antibody [EP1332Y] - Loading Control (ab52866) were incubated overnight at 4°C at 1 in 200 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.
All lanes:
Anti-14-3-3 sigma/SFN antibody [1433S01] (<a href='/en-us/products/unavailable/14-3-3-sigmasfn-antibody-1433s01-ab14123'>ab14123</a>) at 1/200 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
SFN knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human SFN (14-3-3 sigma) knockout HeLa cell line (ab261779)
Lane 3:
A431 cell lysate at 20 µg
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- Sanger seq
Unknown
Sanger Sequencing - Human SFN (14-3-3 sigma) knockout HeLa cell line (AB261779)
Allele-1 : 2 bp deletion in exon 1.
- Sanger seq
Unknown
Sanger Sequencing - Human SFN (14-3-3 sigma) knockout HeLa cell line (AB261779)
Allele-2 : 1 bp deletion in exon 1.
Reactivity data
Product details
Recommended control: Human wild-type HeLa cell line (ab255928). 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 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
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
14-3-3 sigma contributes to cell cycle arrest and apoptosis. As part of a complex it interacts with other proteins to ensure proper cell division and prevent abnormal growth. It binds to phosphorylated serine/threonine residues on its target proteins influencing their activity and function. By doing this it exerts control over cell cycle checkpoints and influences DNA damage response mechanisms.
Pathways
14-3-3 sigma interacts with several pathways that are critical for maintaining cell stability and response to stress. It participates in the PI3K/AKT pathway working closely with proteins like AKT1 to facilitate cell survival and growth suppression under stress conditions. Its involvement in this pathway ties it to the regulation of cell proliferation and survival interconnecting with broader cellular networks.
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
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![Western blot - Anti-14-3-3 sigma/SFN antibody [EPR5873(2)] (AB193667)](https://content.abcam.com/products/images/14-3-3-sigma-sfn-antibody-epr58732-ab193667--western-blot-img66132.jpg)
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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