LMNB1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 2 bp deletion in exon 1 and 2 bp insertion in exon 1.
ADLD, LMN, LMN 2, LMNB, LMNB1_HUMAN, Lamin-B1, MGC111419, OTTHUMP00000159218
LMNB1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 2 bp deletion in exon 1 and 2 bp insertion in exon 1.
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, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.
Lamin B1 also known as LMNB1 is a protein that plays an important role in the structure of the nuclear lamina a dense fibrillar network inside the nucleus of cells. This protein belongs to the lamin family and has an approximate molecular weight of 66 kDa. Lamin B1 is expressed in nearly all types of cells acting as a structural component by interacting with chromatin and anchoring the nuclear envelope. It contributes to regulating nuclear stability and mechanical support which is essential for proper cell function.
Lamin B1 serves many important functions by providing structural integrity and regulating gene expression through chromatin organization. Part of the nuclear lamina complex Lamin B1 interacts with other lamin proteins such as Lamin A and Lamin C forming a mesh-like structure that supports the nuclear envelope. This protein also affects cellular processes such as DNA replication RNA transcription and cell division indicating its broad impact on various cellular activities.
Lamin B1 plays an important role in gene expression regulation and cell cycle control. It interacts with pathways involved in chromatin remodeling influencing the access of transcription factors to DNA which affects gene expression profiles. Lamin B1 associates with related proteins such as Lamin A/C and other nuclear matrix proteins to maintain cellular architecture stability and normal cell function. This organization is vital to ensure the accuracy of cellular processes and prevent anomalies during cell division.
Mutations or altered expression of Lamin B1 have been associated with several pathological conditions. One noteworthy disorder is autosomal dominant adult-onset leukodystrophy where Lamin B1 expression levels are abnormal. This protein also connects to multiple sclerosis through its role in maintaining nuclear architecture. Additionally changes in Lamin B1 expression levels can influence pathways involving Lamin A/C and potentially lead to other laminopathies highlighting its influence in maintaining cellular health and contributing to disease development.
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Anti-Lamin B1 antibody [EPR22165-121] - Nuclear Envelope Marker ab229025 was shown to react with Lamin B1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab255404 (knockout cell lysate Human LMNB1 (Lamin B1) knockout HeLa cell lysate ab263825) was used. Wild-type HeLa and LMNB1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. Anti-Lamin B1 antibody [EPR22165-121] - Nuclear Envelope Marker ab229025 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) overnight at 4° at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (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-Lamin B1 antibody [EPR22165-121] - Nuclear Envelope Marker (Anti-Lamin B1 antibody [EPR22165-121] - Nuclear Envelope Marker ab229025) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: LMNB1 knockout HeLa cell lysate at 20 µg
Lane 2: Western blot - Human LMNB1 (Lamin B1) knockout HeLa cell line (ab255404)
Performed under reducing conditions.
Predicted band size: 66 kDa
Observed band size: 66-70 kDa
Anti-Lamin B1 antibody [EPR8985(B)] - Nuclear Envelope Marker ab133741 was shown to react with LMNB1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab255404 (knockout cell lysate Human LMNB1 (Lamin B1) knockout HeLa cell lysate ab263825) was used. Wild-type HeLa and LMNB1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. Anti-Lamin B1 antibody [EPR8985(B)] - Nuclear Envelope Marker ab133741 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) overnight at 4° at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (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-Lamin B1 antibody [EPR8985(B)] - Nuclear Envelope Marker (Anti-Lamin B1 antibody [EPR8985(B)] - Nuclear Envelope Marker ab133741) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: LMNB1 knockout HeLa cell lysate at 20 µg
Lane 2: Western blot - Human LMNB1 (Lamin B1) knockout HeLa cell line (ab255404)
Performed under reducing conditions.
Predicted band size: 66 kDa
Observed band size: 66-70 kDa
Allele-2: 2 bp insertion in exon 1.
Allele-1: 2 bp deletion in exon 1.
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