Human SNCA (Alpha-synuclein) knockout U-87 MG cell line
- Advanced Validation
- What is this?
Be the first to review this product! Submit a review
|
(0 Publication)
Human SNCA (Alpha-synuclein) knockout U-87 MG cell line available to order. Recommended control: Human wild-type U-87 MG cell line (ab278079).
View Alternative Names
Alpha-synuclein, NACP, Non-A beta component of AD amyloid, Non-A4 component of amyloid precursor, PARK 1, PARK 4, PD-1, SNCA, SYUA_HUMAN, Synuclein alpha, Synuclein alpha 140
- WB
Lab
Western blot - Human SNCA (Alpha-synuclein) knockout U-87 MG cell line (AB282333)
False colour image of Western blot : Anti-Alpha-synuclein antibody [EPR20535] staining at 1/1000 dilution shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution shown in red. In Western blot ab212184 was shown to bind specifically to Alpha-synuclein. A band was observed at 16 kDa in wild-type U-87 MG cell lysates with no signal observed at this size in SNCA knockout cell line ab282333 (knockout cell lysate ab283006). To generate this image wild-type and SNCA knockout U-87 MG 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°. 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 (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
All lanes:
Western blot - Anti-Alpha-synuclein antibody [EPR20535] (<a href='/en-us/products/primary-antibodies/alpha-synuclein-antibody-epr20535-ab212184'>ab212184</a>) at 1/1000 dilution
Lane 1:
Wild-type U-87 MG cell lysate at 20 µg
Lane 2:
SNCA knockout U-87 MG cell lysate at 20 µg
Lane 2:
Western blot - Human SNCA (Alpha-synuclein) knockout U-87 MG cell line (ab282333)
Predicted band size: 14 kDa
Observed band size: 16 kDa
false
- Sanger seq
Lab
Sanger Sequencing - Human SNCA (Alpha-synuclein) knockout U-87 MG cell line (AB282333)
42 bp deletion in exon 2 and intron 2-3, destroy donor splicing site and lead to stop codon
Reactivity data
Product details
Recommended control: Human wild-type U-87 MG cell line (ab278079). 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 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.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Zygosity
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
EMEM + 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
Alpha-synuclein manages several cellular processes most notably in neurotransmitter release and vesicle recycling. It does not work alone; it often forms complexes with other proteins like synaptobrevin-2 of the SNARE complex. The formation of these complexes allows for the proper functioning of synaptic vesicles. Alteration in alpha-synuclein's normal function through aggregation disrupts these processes which underlies many neural complications.
Pathways
Many neuronal functions depend on alpha-synuclein's participation in synaptic vesicle pathways. Key pathways include the dopamine metabolic pathway where alpha-synuclein regulates dopamine release. The protein interacts with other synaptic proteins like the SNARE proteins potentially affecting protein kinase pathways. Abnormal aggregation impacts these pathways which can propagate cellular stress and contribute to cellular degeneration.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
Product protocols
- Visit the General protocols
- Visit the Troubleshooting
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