Human LMNA (Lamin A) knockout HeLa cell line
- Advanced Validation
- What is this?
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(0 Publication)
- WB
Supplier Data
Western blot - Human LMNA (Lamin A) knockout HeLa cell line (AB261787)
Lanes 1-2 : Merged signal (red and green). Green - ab232730 observed at 7075 kDa. Red - loading control ab181602 observed at 37 kDa.
ab232730 Anti-Lamin A + C antibody [WL4G10] was shown to specifically react with Lamin A + C in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261787 (knockout cell lysate ab256979) was used. Wild-type and Lamin A + C knockout samples were subjected to SDS-PAGE. ab232730 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) were incubated overnight at 4° at 1 in 1000 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 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-Lamin A + Lamin C antibody [WL4G10] (<a href='/en-us/products/primary-antibodies/lamin-a-lamin-c-antibody-wl4g10-ab232730'>ab232730</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
Lamin A + C knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human LMNA (Lamin A) knockout HeLa cell line (ab261787)
Secondary
All lanes:
Western blot - Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-800cw-preadsorbed-ab216772'>ab216772</a>) at 1/20000 dilution
Predicted band size: 74 kDa
Observed band size: 70 kDa,75 kDa
false
- WB
Lab
Western blot - Human LMNA (Lamin A) knockout HeLa cell line (AB261787)
Lanes 1-2 : Merged signal (red and green). Green - ab238303 observed at 74 kDa. Red - loading control ab181602 observed at 37 kDa.
ab238303 Anti-Lamin A + C antibody [4C11] was shown to specifically react with Lamin A + C in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261787 (knockout cell lysate ab256979) was used. Wild-type and Lamin A + C knockout samples were subjected to SDS-PAGE. ab238303 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) were incubated overnight at 4° at 1 μg/ml 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 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-Lamin A + Lamin C antibody [4C11] (<a href='/en-us/products/primary-antibodies/lamin-a-lamin-c-antibody-4c11-ab238303'>ab238303</a>) at 1 µg/mL
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
Lamin A + C knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human LMNA (Lamin A) knockout HeLa cell line (ab261787)
Secondary
All lanes:
Western blot - Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-800cw-preadsorbed-ab216772'>ab216772</a>) at 1/20000 dilution
Predicted band size: 74 kDa
Observed band size: 74 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human LMNA (Lamin A) knockout HeLa cell line (AB261787)
Allele-3 : 22 bp deletion in exon 4.
- Sanger seq
Unknown
Sanger Sequencing - Human LMNA (Lamin A) knockout HeLa cell line (AB261787)
Allele-2 : 1 bp deletion in exon 4.
- Sanger seq
Unknown
Sanger Sequencing - Human LMNA (Lamin A) knockout HeLa cell line (AB261787)
Allele-1 : 19 bp deletion in exon 4.
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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
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
The lamin molecule interacts with several nuclear components forming part of the nuclear lamina complex. It anchors chromatin material and regulates DNA replication and repair. Lamin A protein also modulates gene expression through interactions with transcription factors. Beyond structural support it influences cell cycle progression and differentiation impacting cellular mechanotransduction and signaling processes.
Pathways
Lamin A performs critical functions within the cell cycle and apoptotic pathways. Its interactions with the retinoblastoma protein (pRB) and other cyclin-dependent kinases control cell cycle checkpoints and progression. Moreover lamin A connects with proteins involved in signaling pathways like MAPK which relate to stress responses and cellular growth. These interactions highlight its dynamic involvement in maintaining cell health and proliferation.
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
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