Human LIMK1 knockout A549 cell line
- Advanced Validation
- What is this?
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LIMK1 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control available. Knockout. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
View Alternative Names
EC 2.7.1.37, LIM domain containing protein kinase, LIM domain kinase 1, LIM kinase, LIM motif containing protein kinase, LIMK, LIMK1_HUMAN
- WB
Lab
Western blot - Human LIMK1 knockout A549 cell line (AB301040)
Western blot : Anti-LIMK1 antibody [OTI3G3] (ab119084) staining at 1/2000 dilution, shown in green; Rabbit anti-alpha Tubulin antibody [EP1332Y] (ab52866) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab119084 was shown to bind specifically to LIMK1. A band was observed at 73 kDa in wild-type A549 cell lysates with no signal observed at this size in LIMK1 knockout cell line. To generate this image, wild-type and LIMK1 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution.
All lanes:
Western blot - Anti-LIM Kinase 1 antibody [OTI3G3] (<a href='/en-us/products/primary-antibodies/lim-kinase-1-antibody-oti3g3-ab119084'>ab119084</a>) at 1/2000 dilution
Lane 1:
Wild-type A549 cell lysate at 20 µg
Lane 2:
LIMK1 knockout A549 cell lysate at 20 µg
Lane 3:
Wild-type HAP1 cell lysate at 20 µg
Lane 4:
LIMK1 knockout HAP1 cell lysate at 20 µg
Secondary
All lanes:
Goat anti-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution
Observed band size: 73 kDa
false
- NGS
Lab
Next Generation Sequencing - Human LIMK1 knockout A549 cell line (AB301040)
167 bp deletion after Pro 144 (allele 1); 166 bp deletion after Pro 144 (allele 2); 78 bp deletion and 1 bp insertion after Pro 144 (allele 3) of WT protein
Reactivity data
Product details
Although we aim to provide customers with a homozygous clone, feasibility will be dependent on the biology of the protein. Should only heterozygous edits be achieved, you will be notified of the outcome and be asked to confirm whether the cell line is acceptable. All clones will be accompanied with DNA sequencing data, and the mutation description.
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
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.
- Do not allow the cell density to exceed 7x104 cells/cm2.
Culture medium
F-12K + 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 kinase modulates cell shape migration and differentiation by influencing actin filament networks. LIMK1 associates with regulatory complexes and interacts with several proteins to facilitate cytoskeletal reorganization. It plays an essential part in neural development and synaptic plasticity affecting learning and memory. By phosphorylating key actin-binding proteins like cofilin LIMK1 alters actin dynamics essential for various cellular functions.
Pathways
LIM Kinase 1 influences the RhoA/ROCK and Rac1/Pak pathways both critical for cytoskeletal rearrangements. Within these pathways LIMK1 interacts closely with proteins such as cofilin and is influenced by upstream activators like Rho GTPases. These pathways link it to a broader network of signaling that controls cell movement and stability important for cellular responses in various environmental contexts.
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
Target data
Complementary Products
![Western blot - Anti-LIM Kinase 1 antibody [EPR912] (AB108507)](https://content.abcam.com/products/images/lim-kinase-1-antibody-epr912-ab108507--western-blot-img131378.jpg)
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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