Anti-Alpha-synuclein antibody [EPR20535] ab212184 is a rabbit monoclonal antibody that is used in Alpha-synuclein western blotting and IHC. Suitable for human, mouse and rat samples.
- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Specificity confirmed with SNCA knockout cell line validation
- Antibody clone EPR20535 is cited in over 30 publications
IgG
Rabbit
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Monoclonal
WB | IHC-P | IHC-Fr | IP | |
---|---|---|---|---|
Human | Tested | Tested | Expected | Expected |
Mouse | Tested | Tested | Tested | Tested |
Rat | Tested | Tested | Expected | Expected |
Recombinant full length protein | Expected | Expected | Expected | Expected |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info 1/1000 | Notes - |
Species Mouse | Dilution info 1/1000 | Notes - |
Species Human | Dilution info 1/1000 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Recombinant full length protein | Dilution info 1/1000 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/16000 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species Rat | Dilution info 1/16000 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species Human | Dilution info 1/16000 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Recombinant full length protein | Dilution info 1/16000 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/100 | Notes Antigen retrieval: Heated citrate solution (10mM citrate pH 6.0 + 0.05% Tween-20). |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info - | Notes - |
Species Human | Dilution info - | Notes - |
Species Recombinant full length protein | Dilution info 1/100 | Notes Antigen retrieval: Heated citrate solution (10mM citrate pH 6.0 + 0.05% Tween-20). |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/30 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info - | Notes - |
Species Human | Dilution info - | Notes - |
Species Recombinant full length protein | Dilution info 1/30 | Notes - |
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Neuronal protein that plays several roles in synaptic activity such as regulation of synaptic vesicle trafficking and subsequent neurotransmitter release (PubMed:20798282, PubMed:26442590, PubMed:28288128, PubMed:30404828). Participates as a monomer in synaptic vesicle exocytosis by enhancing vesicle priming, fusion and dilation of exocytotic fusion pores (PubMed:28288128, PubMed:30404828). Mechanistically, acts by increasing local Ca(2+) release from microdomains which is essential for the enhancement of ATP-induced exocytosis (PubMed:30404828). Acts also as a molecular chaperone in its multimeric membrane-bound state, assisting in the folding of synaptic fusion components called SNAREs (Soluble NSF Attachment Protein REceptors) at presynaptic plasma membrane in conjunction with cysteine string protein-alpha/DNAJC5 (PubMed:20798282). This chaperone activity is important to sustain normal SNARE-complex assembly during aging (PubMed:20798282). Also plays a role in the regulation of the dopamine neurotransmission by associating with the dopamine transporter (DAT1) and thereby modulating its activity (PubMed:26442590).
NACP, PARK1, PARK1, NACP, SNCA, Alpha-synuclein, Non-A beta component of AD amyloid, Non-A4 component of amyloid precursor, NACP
Anti-Alpha-synuclein antibody [EPR20535] ab212184 is a rabbit monoclonal antibody that is used in Alpha-synuclein western blotting and IHC. Suitable for human, mouse and rat samples.
- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Specificity confirmed with SNCA knockout cell line validation
- Antibody clone EPR20535 is cited in over 30 publications
IgG
Rabbit
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Monoclonal
EPR20535
Affinity purification Protein A
Blue Ice
1-2 weeks
+4°C
-20°C
Upon delivery aliquot
Avoid freeze / thaw cycle
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
This product is a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
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.
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.
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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We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
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False colour image of Western blot: Anti-Alpha-synuclein antibody [EPR20535] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control 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 Human SNCA (Alpha-synuclein) knockout U-87 MG cell line ab282333 (knockout cell lysate Human SNCA (Alpha-synuclein) knockout U-87 MG 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.
All lanes: Western blot - Anti-Alpha-synuclein antibody [EPR20535] (ab212184) 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
Performed under reducing conditions.
Predicted band size: 14 kDa
Observed band size: 16 kDa
Lane 1: Wild type HAP1 whole cell lysate (20 μg)
Lane 2: SNCA (alpha Synuclein) knockout HAP1 whole cell lysate (20 μg)
Lane 3: Human brain whole tissue lysate (20 μg)
Lane 4: SH-SY5Y whole cell lysate (20 μg)
Lanes 1 - 4: Merged signal (red and green). Green - ab212184 observed at 14 kDa. Red - loading control, Anti-GAPDH antibody [mAbcam 9484] - Loading Control ab9484, observed at 37 kDa.
ab212184 was shown to specifically react with SNCA (alpha Synuclein) in wild type cells as signal was lost in SNCA (alpha Synuclein) knockout cells. Wild-type and SNCA (alpha Synuclein) knockout samples were subjected to SDS-PAGE. ab212184 and Anti-GAPDH antibody [mAbcam 9484] - Loading Control ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at a 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-Alpha-synuclein antibody [EPR20535] (ab212184)
Predicted band size: 14 kDa
Immunohistochemical analysis of paraffin-embedded human cerebral cortex tissue labeling Alpha-synuclein with ab212184 at 1/16000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Cytoplasmic staining on human cerebral cortex [PMID: 22112368].
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen mouse hippocampus tissue labeling Alpha-synuclein with ab212184 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (green).
Cytoplasmic staining on mouse hippocampus (PMID: 22112368).
The nuclear counterstain is DAPI (blue).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/1000 dilution.
Blocking/Dilution buffer: 5% NFDM/TBST.
The observed molecular weight is consistent with the literature (PMID: 11739566).
All lanes: Western blot - Anti-Alpha-synuclein antibody [EPR20535] (ab212184) at 1/1000 dilution
Lane 1: Human cerebellum lysate at 20 µg
Lane 2: Human brain lysate at 20 µg
Lane 3: Mouse brain lysate at 20 µg
Lane 4: Rat brain lysate at 20 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 14 kDa
Observed band size: 18 kDa
Exposure time: 5s
Immunohistochemical analysis of paraffin-embedded human glioma tissue labeling Alpha-synuclein with ab212184 at 1/16000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Cytoplasmic staining on human glioma [PMID: 22112368].
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Blocking/Dilution buffer: 5% NFDM/TBST.
Human Alpha-synuclein recombinant protein contain aa1-140 with His-tag. Human Beta-synuclein recombinant protein contain aa1-134 with His-tag.
All lanes: Western blot - Anti-Alpha-synuclein antibody [EPR20535] (ab212184) at 1/1000 dilution
Lane 1: Human Alpha-synuclein recombinant protein at 0.01 µg
Lane 2: Human Beta-synuclein recombinant protein at 0.01 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 14 kDa
Observed band size: 18 kDa
Exposure time: 8s
Alpha-synuclein was immunoprecipitated from 0.35 mg of mouse brain lysate with ab212184 at 1/30 dilution.
Western blot was performed from the immunoprecipitate using ab212184 at 1/1000 dilution.
VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/1000 dilution
Lane 1: Mouse brainlysate 10ug (Input).
Lane 2: ab212184 IP in mouse brain lysate.
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab212184 in mouse brain lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 1 second.
All lanes: Immunoprecipitation - Anti-Alpha-synuclein antibody [EPR20535] (ab212184)
Predicted band size: 14 kDa
Immunohistochemical analysis of paraffin-embedded mouse cerebral cortex tissue labeling Alpha-synuclein with ab212184 at 1/16000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Cytoplasmic staining on mouse cerebral cortex [PMID: 22112368].
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemical analysis of paraffin-embedded rat cerebral cortex tissue labeling Alpha-synuclein with ab212184 at 1/16000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Cytoplasmic staining on rat cerebral cortex [PMID: 22112368].
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling Alpha-synuclein with ab212184 at 1/16000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Negative control: No staining on human kidney. [PMID: 14997013].
Counter stained with Hematoxylin.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
In Western blot, Anti-Alpha-synuclein antibody [EPR20535] (ab212184) staining at 1/1000 dilution.
In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (Anti-6X His tag® antibody [EPR20547] - ChIP Grade ab213204) staining at 1/5000 dilution.
All lanes: Western blot - Anti-Alpha-synuclein (phospho Y39) antibody [EPR25118-5] (Anti-Alpha-synuclein (phospho Y39) antibody [EPR25118-5] ab317366) at 1/1000 dilution
Lane 1: 293T (human embryonic kidney epithelial cell) transfected with humanTyrosine-protein kinase ABL1 expression vector and a human Alpha-synuclein protein (WT) expression vector containing a myc-His-tag® treated with 1 mM pervanadate for 30 minutes whole cell lysate at 4 µg
Lane 2: 293T transfected with human Tyrosine-protein kinase ABL1 expression vector and a human Alpha-synuclein (mutant Y39) protein expression vector containing a myc-His-tag® treated with 1 mM pervanadate for 30 minutes whole cell lysate at 4 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 25 kDa
Exposure time: 10s
In Western blot, Anti-Alpha-synuclein antibody [EPR20535] (ab212184) staining at 1/1000 dilution.
All lanes: Western blot - Anti-Alpha-synuclein (phospho Y39) antibody [EPR25118-5] (Anti-Alpha-synuclein (phospho Y39) antibody [EPR25118-5] ab317366) at 1/1000 dilution
Lane 1: 293T (human embryonic kidney epithelial cell) transfected with human Tyrosine-protein kinase ABL1 expression vector and a human Alpha-synuclein protein expression vector containi a myc-His-tag® treated with 1 mM pervanadate for 30 minutes whole cell lysate (untreated membrane) at 4 µg
Lane 2: 293T transfected with human Tyrosine-protein kinase ABL1 expression vector and a human Alpha-synuclein protein expression vector containi a myc-His-tag® treated with 1 mM pervanadate for 30 minutes whole cell lysate (alkaline phosphatase treated membrane) at 4 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 25 kDa
Exposure time: 8s
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