SPAG5 KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 19 bp deletion in exon 2 and 2 bp deletion in exon 2 and Insertion of the selection cassette in exon 2.
Images
Key facts
- Cell type
- HEK-293T
- Species or organism
- Human
- Tissue
- Kidney
- Form
- Liquid
- Knockout validation
- Sanger Sequencing
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 19 bp deletion in exon 2 and 2 bp deletion in exon 2 and Insertion of the selection cassette in exon 2
Alternative names
Astrin, Deepest, MAP 126, Mitotic spindle associated protein, Mitotic spindle coiled coil related protein, Mitotic spindle-associated protein p126, SPAG5_HUMAN, Sperm tail protein Spag 5, Sperm-associated antigen 5, hMAP 126
Recommended products
SPAG5 KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 19 bp deletion in exon 2 and 2 bp deletion in exon 2 and Insertion of the selection cassette in exon 2.
Key facts
- Cell type
- HEK-293T
- Species or organism
- Human
- Tissue
- Kidney
- Form
- Liquid
- Knockout validation
- Sanger Sequencing
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 19 bp deletion in exon 2 and 2 bp deletion in exon 2 and Insertion of the selection cassette in exon 2
- Concentration
Properties
- Gene name
- SPAG5
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Sanger 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
Astrin also known as SPAG5 or Deepest is a protein important for cell division. It has a molecular mass of approximately 45 kDa. Researchers find Astrin primarily in the cytosol and preferably expressed in proliferating cells. Astrin plays a significant role in spindle stabilization during mitosis binding to microtubules to ensure proper chromosome segregation. This protein has become a focus of study due to its critical functions during cell cycle progression.
- Biological function summary
Astrin facilitates kinetochore-microtubule attachment participating in the mitotic spindle assembly complex. This complex ensures chromosome alignment and segregation which is critical in cellular replication. Since precise regulation of this process is necessary for genetic stability Astrin's function is indispensable. Its presence influences the structural integrity of dividing cells linking it to the stability of genetic information passed to daughter cells after division.
- Pathways
Astrin operates within the spindle assembly checkpoint and cell cycle pathways. The spindle assembly checkpoint pathway ensures that cells do not progress to anaphase until all chromosomes are properly attached to the spindle apparatus. In synergy with proteins like Ndc80 and KNL1 Astrin maintains attachment integrity. Its interactions with these proteins ensure accurate chromosome segregation protecting against chromosomal instability which can lead to aneuploidy.
- Associated diseases and disorders
Astrin's role connects it to conditions like cancer and developmental disorders. Overexpression of Astrin has associations with tumor progression since its function affects cell proliferation. In the context of cancer proteins like Aurora B also influence Astrin's activity. The misregulation of this pathway can lead to faulty chromosomal alignment and segregation contributing to oncogenesis. Such insights make Astrin a potential biomarker and therapeutic target in cancer research.
Product promise
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Full details and terms and conditions can be found here:
Terms & Conditions.
3 product images
Sanger Sequencing - Human SPAG5 (Astrin/Deepest) knockout HEK-293T cell line (ab266536)
Allele-1: 19 bp deletion in exon 2
Sanger Sequencing - Human SPAG5 (Astrin/Deepest) knockout HEK-293T cell line (ab266536)
Allele-2: 2 bp deletion in exon 2.
Sanger Sequencing - Human SPAG5 (Astrin/Deepest) knockout HEK-293T cell line (ab266536)
Allele-3: Insertion of the selection cassette in exon 2.
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