Human LAMA3 knockout HeLa cell line
- Advanced Validation
- What is this?
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- WB
Lab
Western blot - Human LAMA3 knockout HeLa cell line (AB265663)
Lanes 1 - 3 : Merged signal (red and green). Green - ab151715 observed at 367 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.
ab151715 was shown to react with LAMA3 in wild-type HeLa cells in western blot with loss of signal observed in LAMA3 knockout cell line ab265663 (LAMA3 knockout cell lysate ab257497). Wild-type and LAMA3 knockout HeLa cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab151715 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4° at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-LAMA3 antibody [EPR8266] (<a href='/en-us/products/primary-antibodies/lama3-antibody-epr8266-ab151715'>ab151715</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
LAMA3 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human LAMA3 knockout HeLa cell line (ab265663)
Lane 3:
A431 cell lysate at 20 µg
Predicted band size: 367 kDa
Observed band size: 367 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human LAMA3 knockout HeLa cell line (AB265663)
Homozygous : 1 bp deletion in exon 49.
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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 and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant 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
LAMA3 plays an important role in the formation of stable basement membranes and interacts with other proteins to maintain tissue architecture. As part of the laminin-332 complex alongside beta and gamma laminin chains LAMA3 ensures cell adhesion migration and differentiation processes. Its expression in tissues like skin and mucosal surfaces highlights its function in providing structural support and cellular microenvironment regulation.
Pathways
Laminin subunit alpha-3 is essential in pathways related to cell adhesion and migration such as integrin signaling and epithelial-mesenchymal transition. These pathways are pivotal during wound healing and tissue regeneration. LAMA3 associates with integrins particularly beta-1 and beta-4 integrins facilitating communication between cells and their extracellular environment. The interaction with integrins enables cells to respond to structural changes and external stimuli.
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