Human LAMB1 (Laminin beta 1) knockout HeLa cell line
- Advanced Validation
- What is this?
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- WB
Lab
Western blot - Human LAMB1 (Laminin beta 1) knockout HeLa cell line (AB265414)
Lanes 1- 2 : Merged signal (red and green). Green - ab108536 observed at 200 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.
ab108536 was shown to react with Laminin beta 1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265414 (knockout cell lysate ab257499) was used. Wild-type HeLa and LAMB1 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. ab108536 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) 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-Laminin beta 1 antibody [EPR3189(2)] (<a href='/en-us/products/primary-antibodies/laminin-beta-1-antibody-epr31892-ab108536'>ab108536</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 1:
Western blot - Human LAMB1 (Laminin beta 1) knockout HeLa cell lysate (<a href='/en-us/products/cell-lysates/human-lamb1-laminin-beta-1-knockout-hela-cell-lysate-ab257499'>ab257499</a>)
Lane 2:
LAMB1 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human LAMB1 (Laminin beta 1) knockout HeLa cell line (ab265414)
Predicted band size: 198 kDa
Observed band size: 200 kDa
false
- WB
Lab
Western blot - Human LAMB1 (Laminin beta 1) knockout HeLa cell line (AB265414)
Lanes 1- 2 : Merged signal (red and green). Green - ab109293 observed at 200 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.
ab109293 was shown to react with Laminin beta 1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265414 (knockout cell lysate ab257499) was used. Wild-type HeLa and LAMB1 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. ab109293 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) 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-Laminin beta 1 antibody [EPR3190(2)] (<a href='/en-us/products/primary-antibodies/laminin-beta-1-antibody-epr31902-ab109293'>ab109293</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
LAMB1 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human LAMB1 (Laminin beta 1) knockout HeLa cell line (ab265414)
Predicted band size: 198 kDa
Observed band size: 200 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human LAMB1 (Laminin beta 1) knockout HeLa cell line (AB265414)
Homozygous : Insertion of the selection cassette in exon 3.
Complementary Products
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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
Zygosity
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
This component of the laminin molecule plays a critical role in maintaining the integrity of tissues by organizing cell architecture and mediating cellular signals. Laminin beta 1 is part of a larger laminin protein complex that interacts with a variety of cell surface receptors including integrins and dystroglycan contributing to cell communication and signal transduction. These interactions guide cell orientation and regulate vital processes such as cell movement and growth.
Pathways
Laminin beta 1 participates in the integrin signaling pathway and the PI3K-AKT signaling pathway. It interacts closely with integrin proteins such as integrin alpha-6 and integrin beta-4 during these processes. In integrin signaling the laminin molecule facilitates the binding between cells and their surrounding extracellular matrix. This connection influences cellular responses including survival and motility which are necessary for normal development and wound healing processes.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
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