SMAD5 KO cell line available to order. KO validated by Next Generation Sequencing. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9; X = 1 bp insertion, 2 bp deletion, 4 bp deletion, 20 bp deletion, 11 bp deletion.
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Key facts
- Cell type
- HEK-293
- Species or organism
- Human
- Tissue
- Kidney
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing
- Mutation description
- Knockout achieved by CRISPR/Cas9; X = 1 bp insertion, 2 bp deletion, 4 bp deletion, 20 bp deletion, 11 bp deletion
Alternative names
DKFZp781C1895, DKFZp781O1323, Dwfc, JV5-1, MAD homolog 5, MAD, mothers against decapentaplegic homolog 5, MADH 5, Mothers against DPP homolog 5, Mothers against decapentaplegic homolog 5, MusMLP, SMA and MAD related protein 5, SMAD family member 5, SMAD, mothers against DPP homolog 5, SMAD5_HUMAN, hSmad5, mothers against decapentaplegic, drosophila, homolog of, 5
Recommended products
SMAD5 KO cell line available to order. KO validated by Next Generation Sequencing. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9; X = 1 bp insertion, 2 bp deletion, 4 bp deletion, 20 bp deletion, 11 bp deletion.
Key facts
- Cell type
- HEK-293
- Species or organism
- Human
- Tissue
- Kidney
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing
- Mutation description
- Knockout achieved by CRISPR/Cas9; X = 1 bp insertion, 2 bp deletion, 4 bp deletion, 20 bp deletion, 11 bp deletion
- Concentration
Properties
- Gene name
- SMAD5
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Next Generation Sequencing
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 2 US: 2
- Adherent/suspension
- Adherent
- Gender
- Female
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.
Storage
- Shipped at conditions
- Dry Ice
- Appropriate short-term storage conditions
- -196°C
- Appropriate long-term storage conditions
- -196°C
Notes
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.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
SMAD5 also known as Mothers Against Decapentaplegic Homolog 5 is a protein involved in the signal transduction regulated by the transforming growth factor-beta (TGF-beta) superfamily of cytokines. This protein has a molecular mass of approximately 52 kDa and is ubiquitously expressed across various tissues. Mechanically SMAD5 functions as an intracellular mediator that translocates from the cytoplasm to the nucleus upon activation to regulate transcription. It participates actively in transmitting signals from surface receptors into the nucleus influencing gene expression.
- 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.
- Associated diseases and disorders
SMAD5's function has links to abnormal bone and cartilage development such as in fibrodysplasia ossificans progressiva (FOP) and pulmonary hypertension (PH). These conditions occur due to dysregulation within the BMP signaling pathway. For example mutations in SMAD5 or its regulatory proteins can cause inappropriate signal propagation leading to ossification in FOP or altered vascular remodeling in PH. The interplay between SMAD5 and SMAD4 another key player in these pathways highlights the complex nature of SMAD-related pathologies.
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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
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