Human LMNB1 (Lamin B1) knockout HeLa cell line
- Advanced Validation
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LMNB1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 2 bp deletion in exon 1 and 2 bp insertion in exon 1. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.
View Alternative Names
ADLD, LMN, LMN 2, LMNB, LMNB1_HUMAN, Lamin-B1, MGC111419, OTTHUMP00000159218
- WB
Lab
Western blot - Human LMNB1 (Lamin B1) knockout HeLa cell line (AB255404)
Lanes 1- 2 : Merged signal (red and green). Green - ab229025 observed at 66-70 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
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 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. ab229025 and 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 (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 (ab255404)
Predicted band size: 66 kDa
Observed band size: 66-70 kDa
false
- WB
Lab
Western blot - Human LMNB1 (Lamin B1) knockout HeLa cell line (AB255404)
Lanes 1- 2 : Merged signal (red and green). Green - ab133741 observed at 66-70 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
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 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. ab133741 and 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 (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 (ab255404)
Predicted band size: 66 kDa
Observed band size: 66-70 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human LMNB1 (Lamin B1) knockout HeLa cell line (AB255404)
Allele-1 : 2 bp deletion in exon 1.
- Sanger seq
Unknown
Sanger Sequencing - Human LMNB1 (Lamin B1) knockout HeLa cell line (AB255404)
Allele-2 : 2 bp insertion in exon 1.
Reactivity data
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.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
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.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
Complementary Products
Primary Antibodies
AB16048
Anti-Lamin B1 antibody - Nuclear Envelope Marker
BOND RX™ Validated|KO Validated|Lab Essentials
- 5
- 110
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com