Human SMAD5 knockout HEK-293 cell line
Be the first to review this product! Submit a review
|
(0 Publication)
- NGS
Supplier Data
Next Generation Sequencing - Human SMAD5 knockout HEK-293 cell line (AB269470)
4 bp deletion after Lys15 (allele 1), 1 bp insertion after Arg16 (allele 2), and 2 bp deletion after Lys15 (allele 3) of the WT protein
Complementary Products
Primary Antibodies
AB40771
20ul selling size|KO Validated|RabMAb|Recombinant
![Flow Cytometry (Intracellular) - Anti-SMAD5 antibody [EP619Y] (AB40771)](https://content.abcam.com/products/images/smad5-antibody-ep619y-ab40771--flow-cytometry-intracellular-img57277.jpg)
- 5
- 4
Reactivity data
Product details
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, 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
SMAD5 plays a central role in the bone morphogenetic protein (BMP) signaling pathway which is critical for embryonic development and tissue homeostasis. It forms a complex with receptor-regulated SMADs (R-SMADs) and co-SMAD (SMAD4) upon phosphorylation by BMP type I receptors. This complex then moves to the nucleus where it regulates expression of target genes. Through this mechanism SMAD5 impacts processes such as bone formation differentiation and cellular differentiation.
Pathways
Several key cellular signaling processes involve SMAD5. Primarily it contributes to the BMP pathway which is integral to skeletal development and repair. SMAD5 interacts with other SMAD proteins like SMAD1 and SMAD8 which all act downstream of the BMP receptors. Additionally SMAD5 participates in crosstalk with the TGF-beta signaling pathway enabling the fine-tuning of cellular responses to a variety of external cues. This connection places SMAD5 within a network of signaling events important for regulating different cell functions and maintaining cellular homeostasis.
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
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