Human SMAD1 knockout A549 cell line
- Advanced Validation
- What is this?
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(0 Publication)
- WB
Lab
Western blot - Human SMAD1 knockout A549 cell line (AB277886)
Western blot : Anti-SMAD1 antibody [EPR5522] (ab126761) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, 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] (<a href='/en-us/products/primary-antibodies/smad1-antibody-epr5522-ab126761'>ab126761</a>) 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 <a href='/en-us/products/cell-lines/human-smad1-knockout-hela-cell-line-ab265400'>ab265400</a> cell lysate at 20 µg
Secondary
All lanes:
Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
false
- WB
Lab
Western blot - Human SMAD1 knockout A549 cell line (AB277886)
Western blot : Anti-SMAD1 antibody [EP565Y] (ab33902) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, 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] (<a href='/en-us/products/primary-antibodies/smad1-antibody-ep565y-ab33902'>ab33902</a>) 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 <a href='/en-us/products/cell-lines/human-smad1-knockout-hela-cell-line-ab265400'>ab265400</a> cell lysate at 20 µg
Secondary
All lanes:
Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
false
- NGS
Lab
Next Generation Sequencing - Human SMAD1 knockout A549 cell line (AB277886)
121 bp deletion after Pro 57 of WT protein
Complementary Products
Primary Antibodies
AB33902
20ul selling size|KO Validated|RabMAb|Recombinant
![Immunocytochemistry/ Immunofluorescence - Anti-Smad1 antibody [EP565Y] (AB33902)](https://content.abcam.com/products/images/smad1-antibody-ep565y-ab33902--immunocytochemistry-immunofluorescence-img114472.jpg)
- 5
- 3
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
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
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