Human SNCG (gamma Synuclein) knockout HeLa cell line
- Advanced Validation
- What is this?
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SNCG KO cell line available to order. KO validated by Immunocytochemistry, Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 1 and Insertion of the selection cassette in exon 1. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
View Alternative Names
Gamma-synuclein, SR, SNCG, Synoretin, PRSN, BCSG1, Breast cancer-specific gene 1 protein, Persyn, PERSYN
- ICC/IF
Lab
Immunocytochemistry/ Immunofluorescence - Human SNCG (gamma Synuclein) knockout HeLa cell line (AB265623)
Immunofluorescent analysis of 4% Paraformaldehyde fixed 0.1% Triton X-100 permeabilized wild-type HeLa (human cervical adenocarcinoma epithelial cell) and gamma Synuclein/SNCG knockout HeLa cells (ab265623) labelling gamma Synuclein/SNCG with the target antibody at 1/50 (2.06 μg/ml) followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody as secondary at 1/1000 (2 μg/ml) dilution. Confocal image showing partial nuclear and cytoplasmic staining in wild-type HeLa cells (shown in green), with no staining observed in gamma Synuclein/SNCG knockout HeLa cells. ab7291 Anti-alpha Tubulin mouse monoclonal antibody at 1/1000 (1 μg/ml) followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 (2 μg/ml) dilution was used as a counterstain (shown in magenta). Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
- WB
Lab
Western blot - Human SNCG (gamma Synuclein) knockout HeLa cell line (AB265623)
Lanes 1-3 : Merged signal (red and green). Green - ab52633 observed at 16 kDa. Red - loading control ab8245 observed at 36 kDa.
ab52633 Anti-gamma Synuclein/SNCG antibody [EP1539Y] was shown to specifically react with gamma Synuclein/SNCG antibody in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265623 (knockout cell lysate ab257694) was used. Wild-type and gamma Synuclein/SNCG knockout samples were subjected to SDS-PAGE. ab52633 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-gamma Synuclein/SNCG antibody [EP1539Y] (<a href='/en-us/products/primary-antibodies/gamma-synuclein-sncg-antibody-ep1539y-ab52633'>ab52633</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
SNCG knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human SNCG (gamma Synuclein) knockout HeLa cell line (ab265623)
Lane 3:
SK-BR-3 cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution
Predicted band size: 13 kDa
Observed band size: 16 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human SNCG (gamma Synuclein) knockout HeLa cell line (AB265623)
Allele-1 : 1 bp deletion in exon 1.
- Sanger seq
Unknown
Sanger Sequencing - Human SNCG (gamma Synuclein) knockout HeLa cell line (AB265623)
Allele-2 : Insertion of the selection cassette in exon 1.
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Reactivity data
Product details
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.
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
Gamma synuclein plays a role in modulating neuronal plasticity and may participate in synaptic regulation. It is not known to be a part of a larger protein complex but it interacts with various other proteins influencing neuron differentiation. Although its precise biological functions are not fully characterized studies suggest that gamma synuclein could impact cellular processes like cytoskeletal dynamics and membrane trafficking.
Pathways
The protein gamma synuclein is involved in cellular stress response and neurodegeneration pathways. It is closely related to proteins like alpha synuclein with which it shares structural and functional features. Pathways involving these proteins can influence several processes such as axonal transport and neurotransmitter release. Dysregulation in these pathways often leads to disruptions in cellular homeostasis.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
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. <p>1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.<br>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.<br>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.<br>4. Incubate the culture at 37°C incubator with 5% CO<sub>2</sub>. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.<br>5. Once confluent passage into an appropriate flask at a density of 2x10<sup>4</sup> cells/cm<sup>2</sup>. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.</p>
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.
Target data
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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