Human SMAD3 knockout A549 cell line
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SMAD3 KO cell line available to order. KO validated by. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9; X = 1 bp deletion; Frameshift = 100%.
View Alternative Names
DKFZP586N0721, DKFZp686J10186, HSPC193, HST17436, JV15-2, LDS1C, LDS3, MAD (mothers against decapentaplegic Drosophila) homolog 3, MAD homolog 3, MAD, mothers against decapentaplegic homolog 3, MADH 3, MGC60396, Mad homolog JV15 2, Mad protein homolog, Mothers against DPP homolog 3, Mothers against decapentaplegic homolog 3, SMA and MAD related protein 3, SMAD, SMAD family member 3, SMAD, mothers against DPP homolog 3, SMAD3_HUMAN, hMAD-3, hSMAD3
- WB
Lab
Western blot - Human SMAD3 knockout A549 cell line (AB277888)
All lanes:
Western blot - Anti-Smad3 antibody [EPR19686] - ChIP Grade (<a href='/en-us/products/primary-antibodies/smad3-antibody-epr19686-chip-grade-ab208182'>ab208182</a>) at 1/1000 dilution
Lane 1:
Wild-type A549 cell lysate at 20 µg
Lane 2:
SMAD3 CRISPR-Cas9 edited A549 cell lysate at 20 µg
Secondary
Lanes 1 - 2:
Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution
Lanes 1 - 2:
Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Observed band size: 50 kDa
false
- WB
Lab
Western blot - Human SMAD3 knockout A549 cell line (AB277888)
False colour image of Western blot : Anti-Smad3 antibody [EP568Y] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab40854 was shown to bind specifically to Smad3. A band was observed at 50 kDa in wild-type A549 cell lysates with no signal observed at this size in SMAD3 CRISPR-Cas9 edited cell line ab277888 (CRISPR-Cas9 edited cell lysate None). The band observed in the CRISPR-Cas9 edited lysate lane below 50 kDa is likely to represent a truncated form of Smad3. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and SMAD3 CRISPR-Cas9 edited 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-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes:
Western blot - Anti-Smad3 antibody [EP568Y] (<a href='/en-us/products/primary-antibodies/smad3-antibody-ep568y-ab40854'>ab40854</a>) at 1/1000 dilution
Lane 1:
Wild-type A549 cell lysate at 20 µg
Lane 2:
Western blot - Human SMAD3 knockout A549 cell line (ab277888)
Lane 2:
SMAD3 CRISPR-Cas9 edited A549 cell lysate at 20 µg
Secondary
Lanes 1 - 2:
Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution
Lanes 1 - 2:
Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Predicted band size: 48 kDa
Observed band size: 50 kDa
false
Product details
Recommended control: Human wild-type A549 cell line (ab255450). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.
We will provide viable cells that proliferate on revival.
Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.
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
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
Smad3 acts as a mediator of signal transduction for the TGF-beta (transforming growth factor-beta) superfamily forming a complex with phosphorylated Smad2. This enables it to regulate transcriptional activity influencing cell proliferation differentiation and apoptosis. Smad3 also participates in various cellular processes by interacting with other co-factors and regulatory proteins that aid in fine-tuning its function.
Pathways
Smad3 plays an important role in the TGF-beta signaling pathway where it works closely with Smad4 to propagate the signal. Upon phosphorylation it forms a complex with co-Smad (Smad4) and moves into the nucleus to influence gene expression. Smad3 is also involved in pathways related to oncogenesis and tissue fibrosis indicating its significant role in cellular regulation and response mechanisms.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
Target data
Complementary Products
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AB264958
Human CASP8 (Caspase-8) knockout HeLa cell line
Advanced Validation
- 0
- 0
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