LMNA KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 19 bp deletion in exon 4 and 1 bp deletion in exon 4 and 22 bp deletion in exon 4.
70 kDa lamin, CDDC, EMD2, FPL, FPLD, HGPS, IDC, LAMIN A, LAMIN C, LDP1, LFP, LMN 1, LMN C, LMNA_HUMAN, Lamin-A/C, PRO1, Renal carcinoma antigen NY-REN-32
LMNA KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 19 bp deletion in exon 4 and 1 bp deletion in exon 4 and 22 bp deletion in exon 4.
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.
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.
Lamin A also known as LMNA is a type of lamin protein integral to the nuclear architecture. It has a molecular weight of approximately 70 kDa. This protein localizes mainly to the nuclear lamina a dense fibrillar network inside the inner nuclear membrane found across various cell types. Lamin A serves as a structural scaffold mediating nuclear stability and rigidity. It also plays a role in chromatin organization supporting overall nuclear functions.
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.
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.
Mutations in the lamin A gene associate closely with Hutchinson-Gilford Progeria Syndrome and Emery-Dreifuss Muscular Dystrophy. In these conditions altered lamin A protein affects nuclear shape and chromatin layout disrupting transcriptional regulation. The mutated lamin A protein interacts differently with binding partners like emerin contributing to muscle adipose tissue and multi-system abnormalities. This highlights the critical role lamin A plays in maintaining normal cellular function across life stages.
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Anti-Lamin A + Lamin C antibody [WL4G10] 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 Human LMNA (Lamin A) knockout HeLa cell lysate ab256979) was used. Wild-type and Lamin A + C knockout samples were subjected to SDS-PAGE. Anti-Lamin A + Lamin C antibody [WL4G10] ab232730 and Anti-GAPDH antibody[EPR16891] - Loading Control (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 (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (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] (Anti-Lamin A + Lamin C antibody [WL4G10] ab232730) 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)
All lanes: Western blot - Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 74 kDa
Observed band size: 70 kDa, 75 kDa
Anti-Lamin A + Lamin C antibody [4C11] 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 Human LMNA (Lamin A) knockout HeLa cell lysate ab256979) was used. Wild-type and Lamin A + C knockout samples were subjected to SDS-PAGE. Anti-Lamin A + Lamin C antibody [4C11] ab238303 and Anti-GAPDH antibody[EPR16891] - Loading Control (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 (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (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] (Anti-Lamin A + Lamin C antibody [4C11] ab238303) 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)
All lanes: Western blot - Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 74 kDa
Observed band size: 74 kDa
Allele-1: 19 bp deletion in exon 4.
Allele-3: 22 bp deletion in exon 4.
Allele-2: 1 bp deletion in exon 4.
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