SNAP25 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 34 bp Deletion in Exon 2.
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Key facts
- Cell type
- SH-SY5Y
- Species or organism
- Human
- Tissue
- Bone marrow
- Form
- Liquid
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, Homozygous: 34 bp Deletion in Exon 2
Alternative names
Bdr, CMS18, FLJ23079, HGNC:11132, MGC105414, MGC139754, RIC-4, Resistance to inhibitors of cholinesterase 4 homolog, SEC 9, SNAP, SNAP-25B, SNP25_HUMAN, SP, SUP, Super protein, Synaptosomal associated protein 25kDa, Synaptosomal-associated 25 kDa protein, Synaptosomal-associated protein, Synaptosomal-associated protein 25, Synaptosomal-associated protein, 25-KD, bA416N4.2, dJ1068F16.2
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SNAP25 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 34 bp Deletion in Exon 2.
Key facts
- Cell type
- SH-SY5Y
- Species or organism
- Human
- Tissue
- Bone marrow
- Form
- Liquid
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, Homozygous: 34 bp Deletion in Exon 2
- Disease
- Neuroblastoma
- Concentration
Properties
- Gene name
- SNAP25
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Sanger Sequencing, Western blot
- Zygosity
- Homozygous
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 1 US: 1
- 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.
- These cells grow as a mixture of floating and adherent cells.
- Remove media containing floating cells and recover cells by centrifugation, detach cells using standard methods, combine with floating cells and transfer to a new culture flask.
- A guide seeding density of 2x104 cells/cm2 is recommended.
- Cells should be seeded at a density conducive to cell–cell communication to proliferate. If cells are seeded too sparsely, growth rate is reduced and cell death is high.
- Culture medium
- 1:1 mixture of EMEM and F-12K + 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 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.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
SNAP25 also known as Synaptosomal-associated protein 25 plays a critical role in neurotransmitter release through its involvement in the vesicle fusion process. It is part of the SNARE complex and is essential for the docking and fusion of synaptic vesicles with the presynaptic membrane. The protein is approximately 25 kDa in mass and is expressed abundantly in neurons within the central nervous system. Among its various synonyms you may encounter 'protein SNAP' in some literature.
- Biological function summary
SNARE proteins mediate the fusion of transport vesicles with their target membranes. SNAP25 as part of this complex facilitates synaptic-vesicle exocytosis by forming a tight complex with syntaxin and VAMP (also called synaptobrevin). This interaction promotes the merging of vesicle and plasma membranes enabling efficient neurotransmitter release into the synaptic cleft. SNAP25 undergoes dynamic regulation and modification which is important for precise synaptic function.
- Pathways
One observes SNAP25's integration in the process of neurotransmitter release specifically in the exocytosis pathway. It associates closely with other SNARE proteins such as syntaxin-1 and VAMP-2 embodying a core element in synaptic signaling mechanisms. The proper functioning of SNAP25 within this pathway assures the controlled release of neurotransmitters influencing synaptic plasticity and communication.
- Associated diseases and disorders
SNAP25 exhibits strong associations with conditions such as Attention Deficit Hyperactivity Disorder (ADHD) and epilepsy. Variants and dysregulation in SNAP25 have been linked to problems in synaptic transmission which can contribute to these neurological disorders. The protein also interacts with other SNARE complex proteins such as syntaxin which can potentially mediate its role in disease pathogenesis. Understanding these interactions offers insights into therapeutic targets for neurological disorders involving SNAP25.
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3 product images
Western blot - Human SNAP25 knockout SH-SY5Y cell line (ab280041)
False colour image of Western blot: Anti-SNAP25 antibody [EP3274] staining at 1/1000 dilution, shown in green; loading control Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) staining at 1/20000 dilution, shown in red. In Western blot, Anti-SNAP25 antibody [EP3274] ab108990 was shown to bind specifically to SNAP25. A band was observed at 27 kDa in wild-type SH-SY5Y cell lysates with no signal observed at this size in SNAP25 knockout cell line ab280041 (knockout cell lysate Human SNAP25 knockout SH-SY5Y cell lysate ab280100). To generate this image, wild-type and SNAP25 knockout SH-SY5Y 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 (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.
All lanes: Western blot - Anti-SNAP25 antibody [EP3274] (Anti-SNAP25 antibody [EP3274] ab108990) at 1/1000 dilution
Lane 1: Wild-type SH-SY5Y cell lysate at 20 µg
Lane 2: SNAP25 knockout SH-SY5Y cell lysate at 20 µg
Lane 2: Western blot - Human SNAP25 knockout SH-SY5Y cell line (ab280041)
Lane 3: Neuro-2a cell lysate at 20 µg
Lane 4: K562 cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 23 kDa
Observed band size: 27 kDa
Western blot - Human SNAP25 knockout SH-SY5Y cell line (ab280041)
False colour image of Western blot: Anti-SNAP25 antibody [EPR3275] staining at 1/1000 dilution, shown in green; loading control Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) staining at 1/20000 dilution, shown in red. In Western blot, Anti-SNAP25 antibody [EPR3275] ab109105 was shown to bind specifically to SNAP25. A band was observed at 27 kDa in wild-type SH-SY5Y cell lysates with no signal observed at this size in SNAP25 knockout cell line ab280041 (knockout cell lysate Human SNAP25 knockout SH-SY5Y cell lysate ab280100). To generate this image, wild-type and SNAP25 knockout SH-SY5Y 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 (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.
All lanes: Western blot - Anti-SNAP25 antibody [EPR3275] (Anti-SNAP25 antibody [EPR3275] ab109105) at 1/1000 dilution
Lane 1: Wild-type SH-SY5Y cell lysate at 20 µg
Lane 2: SNAP25 knockout SH-SY5Y cell lysate at 20 µg
Lane 2: Western blot - Human SNAP25 knockout SH-SY5Y cell line (ab280041)
Lane 3: Neuro-2a cell lysate at 20 µg
Lane 4: K562 cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 23 kDa
Observed band size: 27 kDa
Sanger Sequencing - Human SNAP25 knockout SH-SY5Y cell line (ab280041)
Human SNAP25 KO in SH-SY5Y Cells with 34 bp Deletion in Exon 2
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