Human SIRT1 knockout A549 cell line
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SIRT1 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided. Knockout.
View Alternative Names
75SirT1, HST2, HST2, S. cerevisiae, homolog of, NAD dependent protein deacetylase sirtuin 1, NAD-dependent deacetylase sirtuin-1, OTTHUMP00000198111, OTTHUMP00000198112, Regulatory protein SIR2 homolog 1, SIR1_HUMAN, SIR2 like 1, SIR2, S.cerevisiae, homolog-like 1, SIR2-like protein 1, SIR2ALPHA, SIR2L1, SirtT1 75 kDa fragment, Sirtuin (silent mating type information regulation 2 homolog) 1 (S. cerevisiae), Sirtuin 1, Sirtuin type 1, hSIR2, hSIRT1
- WB
Lab
Western blot - Human SIRT1 knockout A549 cell line (AB287763)
Western blot : Anti-SIRT1 antibody [EPR18239] (ab189494) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab189494 was shown to bind specifically to SIRT1. A band was observed at 110 kDa in wild-type A549 cell lysates with no signal observed at this size in SIRT1 knockout cell line. To generate this image, wild-type and SIRT1 knockout A549 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-SIRT1 antibody [EPR18239] (<a href='/en-us/products/primary-antibodies/sirt1-antibody-epr18239-ab189494'>ab189494</a>) at 1/1000 dilution
Lane 1:
Wild-type A549 cell lysate at 20 µg
Lane 2:
SIRT1 knockout A549 cell lysate at 20 µg
Lane 3:
Wild-type HEK-293 cell lysate at 20 µg
Lane 4:
SIRT1 knockout HEK-293 cell lysate at 20 µg
Secondary
All lanes:
Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
false
- WB
Lab
Western blot - Human SIRT1 knockout A549 cell line (AB287763)
Western blot : Anti-SIRT1 antibody [1F3] (ab104833) staining at 1/1000 dilution, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab104833 was shown to bind specifically to SIRT1. A band was observed at 110 kDa in wild-type A549 cell lysates with no signal observed at this size in SIRT1 knockout cell line. To generate this image, wild-type and SIRT1 knockout A549 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-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution.
All lanes:
Western blot - Anti-SIRT1 antibody [1F3] - Nuclear Marker (<a href='/en-us/products/primary-antibodies/sirt1-antibody-1f3-nuclear-marker-ab104833'>ab104833</a>) at 1/1000 dilution
Lane 1:
Wild-type A549 cell lysate at 20 µg
Lane 2:
SIRT1 knockout A549 cell lysate at 20 µg
Lane 3:
Wild-type HEK-293 cell lysate at 20 µg
Lane 4:
SIRT1 knockout HEK-293 cell lysate at 20 µg
Secondary
All lanes:
Goat anti-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution
false
- WB
Lab
Western blot - Human SIRT1 knockout A549 cell line (AB287763)
Western blot : Anti-SIRT1 antibody [E104] (ab32441) staining at 1/20000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab32441 was shown to bind specifically to SIRT1. A band was observed at 110 kDa in wild-type A549 cell lysates with no signal observed at this size in SIRT1 knockout cell line. To generate this image, wild-type and SIRT1 knockout A549 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-SIRT1 antibody [E104] (<a href='/en-us/products/primary-antibodies/sirt1-antibody-e104-ab32441'>ab32441</a>) at 1/20000 dilution
Lane 1:
Wild-type A549 cell lysate at 20 µg
Lane 2:
SIRT1 knockout A549 cell lysate at 20 µg
Lane 3:
Wild-type HEK-293 cell lysate at 20 µg
Lane 4:
SIRT1 knockout HEK-293 cell lysate at 20 µg
Secondary
All lanes:
Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
false
- NGS
Lab
Next Generation Sequencing - Human SIRT1 knockout A549 cell line (AB287763)
113 bp deletion after Ala399 (allele 1); 19 bp deletion after Ala399 (allele 2)
Reactivity data
Product details
Although we aim to provide customers with a homozygous clone, feasibility will be dependent on the biology of the protein. Should only heterozygous edits be achieved, you will be notified of the outcome and be asked to confirm whether the cell line is acceptable. All clones will be accompanied with DNA sequencing data, and the mutation description.
Recommended control: Human wild-type A549 cell line (ab288558). 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.
- Do not allow the cell density to exceed 7x104 cells/cm2.
Culture medium
F-12K + 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
SIRT1 modulates several cellular processes such as gene silencing DNA repair and lifespan extension. SIRT1 participates in complexes with other proteins including histones and transcription factors to influence chromatin structure and gene expression. It acts through deacetylation of target proteins affecting their function and stability. The activity of SIRT1 is also linked to environmental and cellular conditions including caloric intake and oxidative stress.
Pathways
SIRT1 is integral in the regulation of metabolic and longevity pathways. It interacts with the FOXO family proteins and the tumor suppressor protein p53 aiding in response to cellular stress and metabolic demands. The role of SIRT1 in the insulin signaling pathway exemplifies its influence on glucose homeostasis and energy balance. These interactions highlight its importance in metabolic health and aging.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
Target data
Complementary Products
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Primary Antibodies
AB110304
BOND RX™ Validated|KO Validated|Recombinant
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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