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AB263769

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

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SNCA KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon2 and Insertion of the selection cassette in exon2.
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Western blot - Human SNCA (Alpha-synuclein) knockout HEK-293T cell lysate (AB263769)
  • WB

Lab

Western blot - Human SNCA (Alpha-synuclein) knockout HEK-293T cell lysate (AB263769)

Lane 1 : Wild-type HEK-293T cell lysate (20ug)
Lane 2 : SNCA knockout HEK-293T cell lysate (20ug)

Lanes 1- 2 : Merged signal (red and green). Green - ab138501 observed at 18 kDa. Red - loading control ab8245 observed at 37 kDa.

ab138501 Recombinant Anti-Alpha-synuclein antibody [MJFR1] was shown to specifically 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 and SNCA knockout samples 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) were incubated overnight at 4oC at 1 in 10000 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.

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 (<a href='/en-us/products/cell-lines/human-snca-alpha-synuclein-knockout-hek-293t-cell-line-ab255433'>ab255433</a>)

Predicted band size: 14 kDa

Observed band size: 18 kDa

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Sanger Sequencing - Human SNCA (Alpha-synuclein) knockout HEK-293T cell lysate (AB263769)
  • Sanger seq

Unknown

Sanger Sequencing - Human SNCA (Alpha-synuclein) knockout HEK-293T cell lysate (AB263769)

Allele-1 : Insertion of the selection cassette in exon2; Allele-2 : 1 bp insertion in exon2

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon2 and Insertion of the selection cassette in exon2.

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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)

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Cell Lines & Lysates

AB255433

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

Advanced Validation

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

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Proteins & Peptides

AB254309

Recombinant human Alpha-Synuclein protein filament

Sandwich ELISA - Recombinant human Alpha-Synuclein protein filament (AB254309)

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  • 0
Secondary Antibodies

AB150077

Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488)

Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (AB150077)

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Reactivity data

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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, 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
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
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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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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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