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AB255433

Human SNCA (Alpha-synuclein) knockout HEK-293T cell line

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(1 Publication)

SNCA KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 2 and Insertion of the selection cassette in exon 2. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
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Western blot - Human SNCA (Alpha-synuclein) knockout HEK-293T cell line (AB255433)
  • WB

Lab

Western blot - Human SNCA (Alpha-synuclein) knockout HEK-293T cell line (AB255433)

** Lanes 1- 2 : ** Merged signal (red and green). Green - ab138501 observed at 18 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa. ab138501 was shown to react with SNCA in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab255433 (knockout cell lysate ab263769) was used. Wild-type HEK-293T and SNCA knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab138501 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 10000 dilution and a 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.

Lanes 1 - 2:

Western blot - Anti-Alpha-synuclein antibody [MJFR1] (<a href='/en-us/products/primary-antibodies/alpha-synuclein-antibody-mjfr1-ab138501'>ab138501</a>) at 1/10000 dilution

Lanes 1 - 2:

Anti-Alpha-synuclein antibody [MJFR1] - BSA and Azide free (<a href='/en-us/products/unavailable/alpha-synuclein-antibody-mjfr1-bsa-and-azide-free-ab156369'>ab156369</a>) at 1/10000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

SNCA knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human SNCA (Alpha-synuclein) knockout HEK-293T cell line (ab255433)

Predicted band size: 14 kDa

Observed band size: 18 kDa

false

Sanger Sequencing - Human SNCA (Alpha-synuclein) knockout HEK-293T cell line (AB255433)
  • Sanger seq

Unknown

Sanger Sequencing - Human SNCA (Alpha-synuclein) knockout HEK-293T cell line (AB255433)

Allele-1 : Insertion of the selection cassette in exon2

Sanger Sequencing - Human SNCA (Alpha-synuclein) knockout HEK-293T cell line (AB255433)
  • Sanger seq

Unknown

Sanger Sequencing - Human SNCA (Alpha-synuclein) knockout HEK-293T cell line (AB255433)

Allele-2 : 1 bp insertion in exon 2.

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Form

Liquid

form

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 2 and Insertion of the selection cassette in exon 2

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Complementary Products

Primary Antibodies

AB138501

Anti-Alpha-synuclein antibody [MJFR1]

BOND RX™ Validated|RabMAb|Recombinant|KO Validated

Western blot - Anti-Alpha-synuclein antibody [MJFR1] (AB138501)

  • 5
  • 10
Cell Lines & Lysates

AB255362

Human APP (Amyloid Precursor Protein) knockout HEK-293T cell line

Advanced Validation

Immunocytochemistry/ Immunofluorescence - Human APP (Amyloid Precursor Protein) knockout HEK-293T cell line (AB255362)

  • 0
  • 0
Cell Lines & Lysates

AB266658

Human CD276 knockout HEK-293T cell line

Advanced Validation

Western blot - Human CD276 knockout HEK-293T cell line (AB266658)

  • 5
  • 2
Cell Lines & Lysates

AB266217

Human DROSHA knockout HEK-293T cell line

Advanced Validation

Western blot - Human DROSHA knockout HEK-293T cell line (AB266217)

  • 0
  • 0
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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
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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.

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What's included?

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Properties and storage information

Gene name
SNCA
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C
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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.

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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

Alpha-synuclein often referred to by alternate names such as SNCA is a protein of around 14 kDa mass. It mainly expresses in the brain particularly in presynaptic nerve terminals. This protein functions mechanically by stabilizing synaptic vesicles and maintaining synaptic function. It exists both in soluble monomer forms and as aggregates in protein filaments. Antibodies like 4D6 and EP1536Y target monomer forms of protein for more detailed studies.
Biological function summary

The alpha-synuclein protein plays critical roles in neuronal activity. It contributes to neurotransmitter release regulation by acting in the formation and plasticity of the presynaptic neuronal network. Alpha-synuclein doesn't usually form parts of large protein complexes but it may associate transiently with membranes and vesicular structures. The protein's monomer form has also been observed in alpha lines and related neuronal processes operating alongside various cellular functions.

Pathways

Synaptic vesicle trafficking and dopamine neurotransmitter release are significant areas involving the alpha-synuclein protein. In these pathways alpha-synuclein interacts with other proteins like synaptophysin and protein monomer monomerizations are intrinsic to these processes. Altered function or aggregation of alpha-synuclein disrupts these pathways influencing broader neurological functions.

Alterations or accumulations of alpha-synuclein are strongly linked to Parkinson's disease and Lewy body dementia. In these conditions alpha-synuclein forms abnormal protein filaments known as Lewy bodies within neurons. These formations disrupt cellular processes and neuron health. Synucleinopathies such as these show connections with proteins like parkin and DJ-1 which also have key roles in these neurodegenerative diseases.
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Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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Product protocols

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Target data

See full target information SNCA
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Publications (1)

Recent publications for all applications. Explore the full list and refine your search

NPJ Parkinson's disease 10:217 PubMed39516469

2024

Novel tools to quantify total, phospho-Ser129 and aggregated alpha-synuclein in the mouse brain.

Applications

Unspecified application

Species

Unspecified reactive species

Benjamin Guy Trist,Courtney Jade Wright,Alejandra Rangel,Louise Cottle,Asheeta Prasad,Nanna Møller Jensen,Hjalte Gram,Nicolas Dzamko,Poul Henning Jensen,Deniz Kirik
View all publications
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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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