LBR KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 2.
DHCR 14B, Integral nuclear envelope inner membrane protein, LBR_HUMAN, LMN 2R, Lamin-B receptor, MGC9041, PHA, PRO0650
LBR KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 2.
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.
The Lamin B Receptor (LBR) also known as L. B. R. L B R or lamin B is a protein located in the inner nuclear membrane. This protein has a molecular mass of approximately 70 kDa. It acts as a receptor for lamin proteins specifically lamin B. The LBR mediates the attachment of heterochromatin and lamins to the inner nuclear membrane playing a structural role in the maintenance of nuclear architecture. It is widely expressed in various tissues notably in cells involved in rapidly proliferating tissues.
The LBR protein serves several critical functions beyond its mechanical duties. It participates in the regulation of chromatin organization and gene expression owing to its interaction with chromatin-related proteins. LBR is part of the nuclear envelope lattice and forms complexes with lamin proteins and heterochromatin. This association supports its role in anchoring chromatin at the nuclear periphery modulating processes such as DNA replication and transcription.
The LBR plays pivotal roles in nuclear envelope assembly and chromatin organization. It holds significance in the cholesterol biosynthesis pathway where it metabolizes sterol intermediates to cholesterol. This function relates LBR to other proteins including lamin A/C which collaborate in maintaining nuclear shape and function. Together they orchestrate various cellular processes important for cell cycle progression and differentiation.
Alterations in LBR function have links to conditions such as Pelger-Huët anomaly and Greenberg skeletal dysplasia. Pelger-Huët anomaly manifests as a benign hematologic condition with defective nuclear shape in neutrophils while Greenberg dysplasia is characterized by skeletal abnormalities. In these contexts LBR connects to lamin B and other nuclear envelope proteins that contribute to the pathophysiological features observed in these disorders.
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Anti-Lamin B Receptor/LBR antibody [E398L] ab32535 was shown to react with Lamin B Receptor/LBR in wild-type HEK-293 cells in western blot. Loss of signal was observed when LBR knockout lysate Human LBR (Lamin B Receptor) knockout HEK-293T cell lysate ab257503 was used. Wild-type and LBR knockout HEK-293 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk before incubation with Anti-Lamin B Receptor/LBR antibody [E398L] ab32535 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4° at a 1 in 500 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (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 B Receptor/LBR antibody [E398L] (Anti-Lamin B Receptor/LBR antibody [E398L] ab32535) at 1/500 dilution
Lane 1: Wild-type HEK-293 cell lysate at 20 µg
Lane 2: Western blot - Human LBR (Lamin B Receptor) knockout HEK-293T cell lysate (Human LBR (Lamin B Receptor) knockout HEK-293T cell lysate ab257503) at 20 µg
Lane 2: Western blot - Human LBR (Lamin B Receptor) knockout HEK-293T cell line (ab266308)
Performed under reducing conditions.
Predicted band size: 71 kDa
Observed band size: 58 kDa
Anti-Lamin B Receptor/LBR antibody [BBmLBR 12.F8] ab232731 was shown to react with Lamin B Receptor (LBR) in wild-type HEK-293 and MEF-1 cells in western blot. Loss of signal was observed when LBR knockout samples were used. Wild-type and LBR knockout (Human LBR (Lamin B Receptor) knockout HEK-293T cell lysate ab257503) HEK-293 cell lysates and wild-type and LBR knockout MEF-1 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk before incubation with Anti-Lamin B Receptor/LBR antibody [BBmLBR 12.F8] ab232731 and Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) overnight at 4° at 1 μg/ml and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (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.
LBR knockout MEF-1 samples were kindly provided by the Brian Burke laboratory A-Star Institute Singapore.
All lanes: Western blot - Anti-Lamin B Receptor/LBR antibody [BBmLBR 12.F8] (Anti-Lamin B Receptor/LBR antibody [BBmLBR 12.F8] ab232731) at 1 µg/mL
Lane 1: Wild-type HEK-293 whole cell lysate at 20 µg
Lane 2: Western blot - Human LBR (Lamin B Receptor) knockout HEK-293T cell lysate (Human LBR (Lamin B Receptor) knockout HEK-293T cell lysate ab257503) at 20 µg
Lane 2: Western blot - Human LBR (Lamin B Receptor) knockout HEK-293T cell line (ab266308)
Lane 3: Wild-type MEF-1 whole cell lysate at 20 µg
Lane 4: LBR knockout MEF-1 whole cell lysate at 20 µg
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: 71 kDa
Observed band size: 58 kDa
Representative images of LBR knockout HEK293T cells, low and high confluency examples (top left and right respectively) and wild-type HEK293T cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using an EVOS M5000 microscope.
Homozygous: 1 bp insertion in exon 2
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