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AB257503

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

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LBR KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 2.
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Western blot - Human LBR (Lamin B Receptor) knockout HEK-293T cell lysate (AB257503)
  • WB

Lab

Western blot - Human LBR (Lamin B Receptor) knockout HEK-293T cell lysate (AB257503)

Lane 1 : Wild-type HEK-293T cell lysate (20 μg)

Lane 2 : LBR knockout HEK-293T cell lysate (20 μg)

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°C 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 (ab257503) at 20 µg

Lane 2:

Western blot - Human LBR (Lamin B Receptor) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-lbr-lamin-b-receptor-knockout-hek-293t-cell-line-ab266308'>ab266308</a>)

Predicted band size: 71 kDa

Observed band size: 58 kDa

false

Sanger Sequencing - Human LBR (Lamin B Receptor) knockout HEK-293T cell lysate (AB257503)
  • Sanger seq

Unknown

Sanger Sequencing - Human LBR (Lamin B Receptor) knockout HEK-293T cell lysate (AB257503)

Homozygous : 1 bp insertion in exon 2

Western blot - Human LBR (Lamin B Receptor) knockout HEK-293T cell lysate (AB257503)
  • WB

Lab

Western blot - Human LBR (Lamin B Receptor) knockout HEK-293T cell lysate (AB257503)

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 HEK-293 cell lysates (ab257503) 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°C 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.

MEF-1 LBR knockout 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 (ab257503) 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

false

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

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

AB32535

Anti-Lamin B Receptor/LBR antibody [E398L]

BOND RX™ Validated|RabMAb|Recombinant|KO Validated|20ul selling size

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lamin B Receptor/LBR antibody [E398L] (AB32535)

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AB290061

Recombinant Human DR5 Protein (Active)

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Biological Activity - Recombinant Human DR5 Protein (Active) (AB290061)

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AB243239

Recombinant mouse Prolactin/PRL protein (Active)

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SDS-PAGE - Recombinant mouse Prolactin/PRL protein (Active) (AB243239)

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "Sanger seq": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" }, "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
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Product details

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.

User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

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?

{ "values": { "1Kit": { "sellingSize": "1 Kit", "publicAssetCode":"ab257503-1Kit", "assetComponentDetails": [ { "size":"1 x 100 µg", "name":"Human LBR knockout HEK293T cell lysate", "number":"AB257503-CMP01", "productcode":"" }, { "size":"1 x 100 µg", "name":"Human wild-type HEK293T cell lysate", "number":"AB257503-CMP02", "productcode":"" } ] } } }
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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
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
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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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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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