Human SNCA (Alpha-synuclein) knockout U-87 MG cell lysate
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- WB
Lab
Western blot - Human SNCA (Alpha-synuclein) knockout U-87 MG cell lysate (AB283006)
Lane 1 : Wild-type U-87 MG cell lysate 20 μg
Lane 2 : SNCA knockout U-87 MG cell lysate 20 μg
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 °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 (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 (<a href='/en-us/products/cell-lines/human-snca-alpha-synuclein-knockout-u-87-mg-cell-line-ab282333'>ab282333</a>)
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 lysate (AB283006)
42 bp deletion in exon 2 and intron 2-3, destroy donor splicing site and lead to stop codon
Complementary Products
Primary Antibodies
AB209538
Anti-Alpha-synuclein aggregate antibody [MJFR-14-6-4-2] - Conformation-Specific
RabMAb|Recombinant
![Immunocytochemistry/ Immunofluorescence - Anti-Alpha-synuclein aggregate antibody [MJFR-14-6-4-2] - Conformation-Specific (AB209538)](https://content.abcam.com/products/images/alpha-synuclein-aggregate-antibody-mjfr-14-6-4-2-conformation-specific-ab209538--immunocytochemistry-immunofluorescence-img66231.jpg)
- 4
- 6
Cell Lines & Lysates
AB255433
Human SNCA (Alpha-synuclein) knockout HEK-293T cell line
Advanced Validation
- 0
- 0
Reactivity data
Product details
Knockout cell lysate achieved by CRISPR/Cas9.
REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.
User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.
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
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
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 promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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