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AB266308

Human LBR (Lamin B Receptor) knockout HEK-293T cell line

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LBR KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 2. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
4 Images
Western blot - Human LBR (Lamin B Receptor) knockout HEK-293T cell line (AB266308)
  • WB

Lab

Western blot - Human LBR (Lamin B Receptor) knockout HEK-293T cell line (AB266308)

Lanes 1 - 4 : Merged signal (red and green). Green - ab232731 observed at 58 kDa. Red - loading control ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) observed at 37kDa.

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 (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 ab232731 and 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 (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (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] (<a href='/en-us/products/primary-antibodies/lamin-b-receptor-lbr-antibody-bbmlbr-12f8-ab232731'>ab232731</a>) 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 (<a href='/en-us/products/cell-lysates/human-lbr-lamin-b-receptor-knockout-hek-293t-cell-lysate-ab257503'>ab257503</a>) at 20 µg

Lane 3:

Wild-type MEF-1 whole cell lysate at 20 µg

Lane 4:

LBR knockout MEF-1 whole cell lysate at 20 µg

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: 71 kDa

Observed band size: 58 kDa

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Western blot - Human LBR (Lamin B Receptor) knockout HEK-293T cell line (AB266308)
  • WB

Lab

Western blot - Human LBR (Lamin B Receptor) knockout HEK-293T cell line (AB266308)

Lanes 1 - 2 : Merged signal (red and green). Green - ab32535 observed at 58 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

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 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 ab32535 and 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 (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (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] (<a href='/en-us/products/primary-antibodies/lamin-b-receptor-lbr-antibody-e398l-ab32535'>ab32535</a>) 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 (<a href='/en-us/products/cell-lysates/human-lbr-lamin-b-receptor-knockout-hek-293t-cell-lysate-ab257503'>ab257503</a>) at 20 µg

Lane 2:

Western blot - Human LBR (Lamin B Receptor) knockout HEK-293T cell line (ab266308)

Predicted band size: 71 kDa

Observed band size: 58 kDa

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Sanger Sequencing - Human LBR (Lamin B Receptor) knockout HEK-293T cell line (AB266308)
  • Sanger seq

Unknown

Sanger Sequencing - Human LBR (Lamin B Receptor) knockout HEK-293T cell line (AB266308)

Homozygous : 1 bp insertion in exon 2

Cell Culture - Human LBR (Lamin B Receptor) knockout HEK-293T cell line (AB266308)
  • Cell Culture

Unknown

Cell Culture - Human LBR (Lamin B Receptor) knockout HEK-293T cell line (AB266308)

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.

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Form

Liquid

form

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 2

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Complementary Products

Primary Antibodies

AB232731

Anti-Lamin B Receptor/LBR antibody [BBmLBR 12.F8]

BOND RX™ Validated|KO Validated

Immunocytochemistry/ Immunofluorescence - Anti-Lamin B Receptor/LBR antibody [BBmLBR 12.F8] (AB232731)

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Cell Lines & Lysates

AB266257

Human VRK1 knockout HEK-293T cell line

Advanced Validation

Western blot - Human VRK1 knockout HEK-293T cell line (AB266257)

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Cell Lines & Lysates

AB266336

Human EMD (Emerin) knockout HEK-293T cell line

Advanced Validation

Western blot - Human EMD (Emerin) knockout HEK-293T cell line (AB266336)

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Cell Lines & Lysates

AB273748

Human BSG (CD147) knockout A549 cell line

Advanced Validation

Flow Cytometry - Human BSG (CD147) knockout A549 cell line (AB273748)

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Reactivity data

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Product details

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.

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What's included?

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Properties and storage information

Gene name
LBR
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Zygosity
Homozygous
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C
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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.

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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

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.
Biological function summary

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.

Pathways

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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Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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Product protocols

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Target data

See full target information LBR
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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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