Human SMN1 knockout A549 cell line
- Advanced Validation
- What is this?
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SMN1 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control available. 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
SMNT, SMN1, Survival motor neuron protein, Component of gems 1, SMN, Gemin-1, SMNC, SMN2
- WB
Lab
Western blot - Human SMN1 knockout A549 cell line (AB301247)
Western blot : Rabbit Monoclonal [EPR4430] to SMN/Gemin 1 ab108424 staining at 1/1000 dilution, shown in green; Mouse anti alpha Tubulin (ab7291) loading control staining at 1/20,000 dilution, shown in magenta.
A band was observed at 32 kDa in Wild-type A549 cell lysates with no signal observed at this size in SMN1 knockout A549 cell line.
To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc 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-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20,000 dilution.
All lanes:
Western blot - Anti-SMN/Gemin 1 antibody [EPR4430] (<a href='/en-us/products/primary-antibodies/smn-gemin-1-antibody-epr4430-ab108424'>ab108424</a>) at 1/1000 dilution
Lane 1:
Wild-type A549 at 20 µg
Lane 2:
SMN1 knockout A549 at 20 µg
Lane 2:
Western blot - Human SMN1 knockout A549 cell line (ab301247) at 20 µg
Lane 3:
HeLa at 20 µg
Lane 4:
PC-3 at 20 µg
Secondary
All lanes:
Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution
Predicted band size: 32 kDa
Observed band size: 32 kDa
false
- WB
Lab
Western blot - Human SMN1 knockout A549 cell line (AB301247)
Western blot : Rabbit Monoclonal [EPR4429] to SMN/Gemin 1 ab108531 staining at 1/1000 dilution, shown in green; Mouse anti alpha Tubulin (ab7291) loading control staining at 1/20,000 dilution, shown in magenta.
A band was observed at 32 kDa in Wild-type A549 cell lysates with no signal observed at this size in SMN1 knockout A549 cell line.
To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc 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-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20,000 dilution.
All lanes:
Western blot - Anti-SMN/Gemin 1 antibody [EPR4429] (<a href='/en-us/products/primary-antibodies/smn-gemin-1-antibody-epr4429-ab108531'>ab108531</a>) at 1/1000 dilution
Lane 1:
Wild-type A549 at 20 µg
Lane 2:
SMN1 knockout A549 at 20 µg
Lane 2:
Western blot - Human SMN1 knockout A549 cell line (ab301247) at 20 µg
Lane 3:
HeLa at 20 µg
Lane 4:
PC-3 at 20 µg
Secondary
All lanes:
Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution
Predicted band size: 32 kDa
Observed band size: 32 kDa
false
- NGS
Supplier Data
Next Generation Sequencing - Human SMN1 knockout A549 cell line (AB301247)
140 bp deletion after Ala111 of the WT protein
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Advanced Validation
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Reactivity data
Product details
Note: Due to high homology the SMN1 knockout strategy may also impact SMN2, although this has not been tested.
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
The SMN protein operates within the SMN-Gemin complex a multi-protein assembly important for snRNP biogenesis and pre-mRNA splicing. This complex coordinates the maturation and transport of snRNPs facilitating the processing of precursor mRNAs in the nucleus. SMN and associated Gemin proteins mediate the assembly of Sm proteins onto snRNA ensuring accurate splicing mechanisms necessary for gene expression.
Pathways
The SMN protein is notably involved in the chaperone-assisted assembly pathway related to snRNP biogenesis. It plays a central role in the spliceosome assembly process working closely with Sm proteins and other RNA-binding proteins. Importantly SMN interacts with the ribonucleoprotein complex influencing mRNA splicing which integrates into the broader cell signaling pathways such as the cholinergic agonist pathways and RNA splicing pathways that regulate neuromuscular communication.
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
Target data
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