SMAD1 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided.
Images
Key facts
- Cell type
- A549
- Species or organism
- Human
- Tissue
- Lung
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing, Western blot
Alternative names
BSP-1, HsMAD1, JV4-1, MAD homolog 1, MAD mothers against decapentaplegic homolog 1, MADH1, MADR1, Mad-related protein 1, Mothers against DPP homolog 1, Mothers against decapentaplegic homolog 1, SMA- AND MAD-RELATED PROTEIN 1, SMAD family member 1, SMAD mothers against DPP homolog 1, SMAD1_HUMAN, TGF beta signaling protein 1, Transforming growth factor-beta-signaling protein 1
Recommended products
SMAD1 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided.
Key facts
- Cell type
- A549
- Species or organism
- Human
- Tissue
- Lung
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing, Western blot
- Disease
- Carcinoma
- Concentration
Properties
- Gene name
- SMAD1
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Next Generation Sequencing, Western blot
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 1 US: 1
- Adherent/suspension
- Adherent
- Gender
- Male
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.
Storage
- Shipped at conditions
- Dry Ice
- Appropriate short-term storage conditions
- -196°C
- Appropriate long-term storage conditions
- -196°C
Notes
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.
Recommended control: Human wild-type A549 cell line (ab288558). 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.
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.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
Smad1 also known as Mothers against decapentaplegic homolog 1 (MADH1) is a protein encoded by the SMAD1 gene in humans. It has a molecular mass of approximately 53 kDa and functions as a receptor-regulated Smad protein. Smad1 primarily transmits signals from the bone morphogenetic proteins (BMPs) a group of growth factors and cytokines. Expression of Smad1 occurs in various tissues including the lung heart and kidney. It plays an important role in mediating signaling pathways within different cellular environments.
- Biological function summary
Smad1 participates in the transmission of BMP signals from the cell surface to the nucleus ensuring transcriptional regulation of target genes. In the presence of BMPs Smad1 forms complexes with Smad4 upon activation. These Smad complexes then translocate to the nucleus where they regulate gene expression. Smad1 influences cellular responses such as proliferation differentiation and apoptosis. In particular it plays a significant role in bone development and osteogenesis.
- Pathways
Smad1 plays an important role within the BMP signaling pathway which is important for early development and tissue homeostasis. Within this pathway BMPs trigger the phosphorylation of Smad1 which then associates with Smad4 to propagate downstream signaling. Another associated pathway includes the TGF-beta signaling pathway where Smads like Smad2 and Smad3 show functional similarities and differences with Smad1. The interactions of Smad1 with related proteins like Smad4 highlight its significant contribution to cellular processes regulated by these pathways.
- Associated diseases and disorders
Smad1 associates with various pathological conditions. For instance aberrant Smad1 signaling has implications in cancer particularly in processes like tumor progression and metastasis. Disruptions in the BMP pathway involving Smad1 and Smad4 can influence the onset and development of disorders such as pulmonary hypertension where inappropriate cell growth and remodeling occur. Understanding Smad1's role in these contexts provides insights into potential therapeutic avenues for treatment and management of these conditions.
Product promise
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Full details and terms and conditions can be found here:
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3 product images
Western blot - Human SMAD1 knockout A549 cell line (ab277886)
Western blot: Anti-SMAD1 antibody [EP565Y] (Anti-Smad1 antibody [EP565Y] ab33902) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-Smad1 antibody [EP565Y] ab33902 was shown to bind specifically to SMAD1. A band was observed at 55-65 kDa in wild-type A549 cell lysates with no signal observed at this size in SMAD1 knockout cell line. To generate this image, wild-type and SMAD1 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 5 % 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-Smad1 antibody [EP565Y] (Anti-Smad1 antibody [EP565Y] ab33902) at 1/1000 dilution
Lane 1: Wild-type A549 cell lysate at 20 µg
Lane 2: SMAD1 knockout A549 cell lysate at 20 µg
Lane 3: Wild-type HeLa ab255929 cell lysate at 20 µg
Lane 4: SMAD1 knockout HeLa Human SMAD1 knockout HeLa cell line ab265400 cell lysate at 20 µg
SecondaryAll lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Western blot - Human SMAD1 knockout A549 cell line (ab277886)
Western blot: Anti-SMAD1 antibody [EPR5522] (Anti-Smad1 antibody [EPR5522] ab126761) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-Smad1 antibody [EPR5522] ab126761 was shown to bind specifically to SMAD1. A band was observed at 52 kDa in wild-type A549 cell lysates with no signal observed at this size in SMAD1 knockout cell line. To generate this image, wild-type and SMAD1 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-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes: Western blot - Anti-Smad1 antibody [EPR5522] (Anti-Smad1 antibody [EPR5522] ab126761) at 1/1000 dilution
Lane 1: Wild-type A549 cell lysate at 20 µg
Lane 2: SMAD1 knockout A549 cell lysate at 20 µg
Lane 3: Wild-type HeLa ab255929 cell lysate at 20 µg
Lane 4: SMAD1 knockout HeLa Human SMAD1 knockout HeLa cell line ab265400 cell lysate at 20 µg
SecondaryAll lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Next Generation Sequencing - Human SMAD1 knockout A549 cell line (ab277886)
121 bp deletion after Pro 57 of WT protein
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