Human SMAD1 knockout HeLa cell line
- Advanced Validation
- What is this?
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(0 Publication)
SMAD1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 3 and 1 bp insertion in exon 3.
View 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
- WB
Lab
Western blot - Human SMAD1 knockout HeLa cell line (AB265400)
Lanes 1- 4 : Merged signal (red and green). Green - ab126761 observed at 52 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab126761 was shown to react with Smad1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265400 (knockout cell lysate ab257686) was used. Wild-type HeLa and SMAD1 knockout HeLa cell lysates were subjected to SDS-PAGE. ab126761 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° at a 1 in 1000 Dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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 HeLa cell lysate at 20 µg
Lane 2:
SMAD1 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human SMAD1 knockout HeLa cell line (ab265400)
Lane 3:
HT1080 cell lysate at 20 µg
Lane 4:
Huvec cell lysate at 20 µg
Predicted band size: 52 kDa
Observed band size: 52 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human SMAD1 knockout HeLa cell line (AB265400)
Allele-2 : 1 bp insertion in exon 3.
- Sanger seq
Unknown
Sanger Sequencing - Human SMAD1 knockout HeLa cell line (AB265400)
Allele-1 : 1 bp deletion in exon 3.
Reactivity data
Product details
Recommended control: Human wild-type HeLa cell line (ab255928). 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, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and 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.
Culture medium
DMEM (High Glucose) + 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: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
Complementary Products
Primary Antibodies
AB33902
20ul selling size|KO Validated|RabMAb|Recombinant
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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