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AB263825

Human LMNB1 (Lamin B1) knockout HeLa cell lysate

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LMNB1 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 2 bp deletion in exon 1 and 2 bp insertion in exon 1.
4 Images
Western blot - Human LMNB1 (Lamin B1) knockout HeLa cell lysate (AB263825)
  • WB

Lab

Western blot - Human LMNB1 (Lamin B1) knockout HeLa cell lysate (AB263825)

Lane 1 : Wild-type HeLa cell lysate (20µg)

Lane 2 : LMNB1 knockout HeLa cell lysate (20µg)

Lanes 1- 2 : Merged signal (red and green). Green - ab229025 observed at 66-70 kDa. Red - loading control ab8245 observed at 37 kDa.

ab229025 Recombinant Anti-Lamin B1 antibody [EPR22165-121] was shown to specifically react with LMNB1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab255404 (knockout cell lysate ab263825) was used. Wild-type and LMNB1 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab229025 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and 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 (<a href='/en-us/products/primary-antibodies/lamin-b1-antibody-epr22165-121-nuclear-envelope-marker-ab229025'>ab229025</a>) 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 (<a href='/en-us/products/cell-lines/human-lmnb1-lamin-b1-knockout-hela-cell-line-ab255404'>ab255404</a>)

Predicted band size: 66 kDa

Observed band size: 66-70 kDa

false

Western blot - Human LMNB1 (Lamin B1) knockout HeLa cell lysate (AB263825)
  • WB

Lab

Western blot - Human LMNB1 (Lamin B1) knockout HeLa cell lysate (AB263825)

Lane 1 : Wild-type HeLa cell lysate (20µg)

Lane 2 : LMNB1 knockout HeLa cell lysate (20µg)

Lanes 1- 2 : Merged signal (red and green). Green - ab133741 observed at 66-70 kDa. Red - loading control ab8245 observed at 37 kDa.

ab133741 Recombinant Anti-Lamin B1 antibody [EPR8985(B)] was shown to specifically react with LMNB1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab255404 (knockout cell lysate ab263825) was used. Wild-type and LMNB1 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab133741 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and 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 (<a href='/en-us/products/primary-antibodies/lamin-b1-antibody-epr8985b-nuclear-envelope-marker-ab133741'>ab133741</a>) 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 (<a href='/en-us/products/cell-lines/human-lmnb1-lamin-b1-knockout-hela-cell-line-ab255404'>ab255404</a>)

Predicted band size: 66 kDa

Observed band size: 66-70 kDa

false

Sanger Sequencing - Human LMNB1 (Lamin B1) knockout HeLa cell lysate (AB263825)
  • Sanger seq

Unknown

Sanger Sequencing - Human LMNB1 (Lamin B1) knockout HeLa cell lysate (AB263825)

Allele-1 : 2 bp deletion in exon 1

Sanger Sequencing - Human LMNB1 (Lamin B1) knockout HeLa cell lysate (AB263825)
  • Sanger seq

Unknown

Sanger Sequencing - Human LMNB1 (Lamin B1) knockout HeLa cell lysate (AB263825)

Allele-2 : 2 bp insertion in exon 1

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 2 bp deletion in exon 1 and 2 bp insertion in exon 1.

Disease

Adenocarcinoma

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Complementary Products

Primary Antibodies

AB16048

Anti-Lamin B1 antibody - Nuclear Envelope Marker

BOND RX™ Validated|KO Validated|Lab Essentials

Western blot - Anti-Lamin B1 antibody - Nuclear Envelope Marker (AB16048)

  • 5
  • 110
Proteins & Peptides

AB217407

Recombinant human Vitronectin/S-Protein (Animal Free)

  • 0
  • 0
Proteins & Peptides

AB198115

Recombinant Human Histone H4 protein

SDS-PAGE - Recombinant Human Histone H4 protein (AB198115)

  • 0
  • 0
Assay Kits

AB252351

Human Lamin B1 ELISA Kit

Recombinant|SimpleStep

Sandwich ELISA - Human Lamin B1 ELISA Kit (AB252351)

  • 0
  • 0
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Reactivity data

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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?

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Properties and storage information

Gene name
LMNB1
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
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.

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.
Biological function summary

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.

Pathways

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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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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