Human SIRT6 knockout HeLa cell line
- Advanced Validation
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SIRT6 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 2 bp deletion in exon 1 and 4 bp deletion in exon 1. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.
View Alternative Names
2810449N18Rik, AI043036, Mono ADP ribosyltransferase sirtuin 6, NAD-dependent protein deacetylase sirtuin-6, Regulatory protein SIR2 homolog, Regulatory protein SIR2 homolog 6, SIR2 like 6, SIR2-like protein 6, SIR2L6, SIR6_HUMAN, Sir2 related protein type 6, Sirtuin (silent mating type information regulation 2 homolog) 6 (S. cerevisiae), Sirtuin 6, Sirtuin type 6
- ICC/IF
Lab
Immunocytochemistry/ Immunofluorescence - Human SIRT6 knockout HeLa cell line (AB265054)
ab289970 staining SIRT6 in wild-type HeLa (human cervix adenocarcinoma epithelial cell line) cells and SIRT6 knockout HeLa (human cervix adenocarcinoma epithelial cell line) cells (ab265054). The cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. The cells were then incubated with ab289970 at 1/1000 dilution and ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) at 1/200 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) secondary antibody at 1/1000 dilution. Nuclear DNA was labelled in blue with DAPI. Confocal image showing nuclear staining in parental Hela cell line and no staining in SIRT6 KO Hela cell line.
- Flow Cyt (Intra)
Lab
Flow Cytometry (Intracellular) - Human SIRT6 knockout HeLa cell line (AB265054)
- WB
Lab
Western blot - Human SIRT6 knockout HeLa cell line (AB265054)
Lanes 1-2 : Merged signal (red and green). Green - ab176345 observed at 45 kDa. Red - loading control ab8245 observed at 37 kDa.
ab176345 Anti-SIRT6 antibody [EPR5079(N)] was shown to specifically react with SIRT6 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265054 (knockout cell lysate ab257673) was used. Wild-type and SIRT6 knockout samples were subjected to SDS-PAGE. ab176345 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° at 1 in 1000 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 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-SIRT6 antibody [EPR5079(N)] (<a href='/en-us/products/primary-antibodies/sirt6-antibody-epr5079n-ab176345'>ab176345</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
SIRT6 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human SIRT6 knockout HeLa cell line (ab265054)
Predicted band size: 39 kDa
Observed band size: 45 kDa
false
- WB
Lab
Western blot - Human SIRT6 knockout HeLa cell line (AB265054)
Lanes 1-2 : Merged signal (red and green). Green - ab191385 observed at 40 kDa. Red - loading control ab7291 observed at 50 kDa.
ab191385 Anti-SIRT6 antibody [EPR18463] was shown to specifically react with SIRT6 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265054 (knockout cell lysate ab257673) was used. Wild-type and SIRT6 knockout samples were subjected to SDS-PAGE. ab191385 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4° at 1 in 1000 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 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-SIRT6 antibody [EPR18463] (<a href='/en-us/products/primary-antibodies/sirt6-antibody-epr18463-ab191385'>ab191385</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
SIRT6 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human SIRT6 knockout HeLa cell line (ab265054)
Predicted band size: 39 kDa
Observed band size: 40 kDa,42 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human SIRT6 knockout HeLa cell line (AB265054)
Allele-2 : 2 bp deletion in exon 1.
- Sanger seq
Unknown
Sanger Sequencing - Human SIRT6 knockout HeLa cell line (AB265054)
Allele-1 : 4 bp deletion in exon 1.
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
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
SIRT6 influences DNA repair metabolism and inflammation. It participates in maintaining genomic stability by promoting base excision repair a critical DNA repair process. Moreover SIRT6 contributes to glucose homeostasis by influencing gluconeogenesis and glycolysis. This protein is not known to be part of any larger protein complexes but it interacts individually with other proteins to exert its biological effects.
Pathways
SIRT6 plays a significant role in two key biological pathways: DNA damage response and metabolism regulation. In the DNA damage response pathway SIRT6 works with other proteins like PARP1 to facilitate DNA repair under stress conditions. In the regulation of metabolism SIRT6 interacts with transcription factors like HIF1α which influences the expression of genes involved in glycolytic metabolism and glucose homeostasis.
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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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