Invitrogen

alpha Synuclein Monoclonal Antibody (Syn 211)

Advanced Verification

This Antibody was verified by Knockout to ensure that the antibody binds to the antigen stated. View Details

3 Published Figures

17 References

View all (51) alpha Synuclein antibodies

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Datasheet

Protocols

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alpha Synuclein Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescent analysis of Synuclein (green) showing staining in the cytoplasm and membrane of U87-MG cells. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a Synuclein monoclonal antibody (Product # MA5-12272) in 3% BSA-PBS at a dilution of 1:100 and incubated overnight at 4 °C in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

alpha Synuclein Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry analysis of Synuclein showing positive staining in the cytoplasm and nucleus of paraffin-treated Human cerebellum tissue (right) compared with a negative control in the absence of primary antibody (left). To expose target proteins, antigen retrieval method was performed using 10mM sodium citrate (pH 6.0) microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a Synuclein monoclonal antibody (Product # MA5-12272) diluted by 3% BSA-PBS at a dilution of 1:100 overnight at 4°C in a humidified chamber. Tissues were washed extensively PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

alpha Synuclein Antibody in Western Blot (WB) — Western blot was performed using Anti-alpha Synuclein Monoclonal Antibody, (Product # MA5-12272) and a ~14 kDa band corresponding to alpha Synuclein was observed in SH-SY5Y and HEK-293 cell lines and not in Hep G2 and A549 cell lines which are reported be expressing low to nil amounts of alpha Synuclein protein. Whole cell extracts (30 µg lysate) of SH-SY5Y (Lane1), HEK-293 (Lane 2), Hep G2 (Lane 3) and A549 (Lane 4) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001) and the membrane was fixed with 0.4% PFA for 30 mins at room temperature. The blot was later washed with 1X PBS twice and probed with the primary antibody (1 µg/mL) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

alpha Synuclein Antibody — Antibody specificity was demonstrated by CRISPR-Cas9 mediated knockout of alpha Synuclein. A loss of signal was observed for alpha Synuclein in the KO cell line compared to control cell line using Anti- alpha Synuclein Monoclonal Antibody (Product # MA5-12272).. {KO}

alpha Synuclein Antibody in Immunohistochemistry (IHC) — Figure 1 Dopamine transporter (DAT) staining in the caudate putamen of PD patients and age-matched controls. ( A ) Representative photomicrograph showing the negative control produced by performing DAT immunolabeling on cerebellum sections. Please note the absence of staining that is indicative of the specificity of the DAT antibody used for this study; ( B , D , E , G , H , G1 , H1 , G2 , G3 ) Representative images showing DAT immunolabeling in the caudate putamen of PD patients and age-matched controls subjects. DAT immunolabeling showed a small dot-like appearance with widespread distribution in the control sections ( B , D , E ). Please note that in spite of the overall decrease of immunolabeling, in the PD brains, the DAT signal was accumulated in big neuropil-like dots ( G , H , G1 , H1 , G2 , arrowheads) and in few neuritic-like structures ( G , G1 , G3 , arrows); ( C , F , I , I1 , L ) Panels are showing alpha-synuclein immunolabeling in the caudate putamen of PD patients and age-matched control subjects. Alpha-synuclein immunolabeling showed a widespread distribution in the grey matter in control sections ( C , F ). Please note the presence of neuropil-like alpha-synuclein-positive dots ( I , I1 , L , arrowheads) and LB-like structures in the PD brains ( L , arrow). Scale bars: ( A - C , G - I ) 100 um, ( D - F , G1 , H1 , I1 ) 50 um, ( G2 , G3 , L ) 20 um.

alpha Synuclein Antibody in Proximity Ligation Assay (PLA) (PLA) — Figure 4 Dopamine transporter (DAT)/alpha-synuclein in situ PLA performed on the caudate putamen of PD patients and control subjects. ( A - C ) Panels showing the negative control, performed by omitting the DAT antibody, of the in situ PLA assayed on the caudate putamen of a control subject. The absence of the PLA-positive signal is indicative of the specificity of the assay; ( D - U ) Representative images showing DAT/alpha-synuclein in situ PLA performed on the caudate putamen of PD patients ( J - U ) and age-matched-controls (control, D - I ). The presence of a PLA positivity as a red fluorescent signal is indicative of the interaction between the two proteins. Please note the marked redistribution of the PLA-positivity in the caudate putamen of PD samples ( L , O , R ) when compared to the control subjects ( F , I ). In particular, in the PD brains, the PLA signal was particularly abundant within big ( O , arrowheads) and small ( L , R , arrowheads) clumps or neuritic-like structures ( U ). Scale bar: 20 mum.

Product Details

Applications

Tested Dilution

Publications

Western Blot (WB)

1 µg/mL

View 6 publications 6 publications

Immunohistochemistry (IHC)

View 4 publications 4 publications

Immunohistochemistry (Paraffin) (IHC (P))

1:200-1:400

Immunocytochemistry (ICC/IF)

1:20-1:200

View 5 publications 5 publications

Immunoprecipitation (IP)

Assay-dependent

in situ PLA (PLA)

View 1 publication 1 publication

Miscellaneous PubMed (Misc)

View 1 publication 1 publication

Product Specifications

Species Reactivity

Human, Rat

Published species

Human, Mouse

Host/Isotype

Mouse / IgG1

Class

Monoclonal

Type

Antibody

Clone

Syn 211

Immunogen

Human recombinant alpha-synuclein

Conjugate

Unconjugated

Form

Liquid

Concentration

0.2 mg/mL

Purification

Protein G

Storage buffer

PBS, pH 7.4, with 0.2% BSA

Contains

0.09% sodium azide

Storage conditions

4° C

RRID

AB_10978319

Product Specific Information

MA5-12272 targets Synuclein in IHC (P), ICC/IF, IP, and WB applications and shows reactivity with Human and Rat samples.

The MA5-12272 immunogen is human recombinant alpha-synuclein.

Target Information

Alpha-Synuclein belongs to the Synuclein family, which includes beta and gamma Synuclein, and is predominantly expressed in neurons, and concentrated at synaptic terminals. Synucleins may be involved in the regulation of dopamine release and transport and the modulation of synaptic vesicle function. Alpha-Synuclein is a small, presynaptic protein that is the major nonamyloid component of the pathological inclusions characteristic of a wide range of neurodegenerative disorders, collectively known as synucleinopathies. Alpha-Synuclein is involved in the formation of SNARE complexes, and most significantly, aggregated alpha-synuclein is one of the major components found in the Lewy bodies that occur in Parkinson's disease (PD) and other neurodegenerative disorders. Mutation of the alpha-synuclein gene is associated with familial forms of PD. Alpha-Synuclein is known to reduce the fibrillization of the microtubule-associated protein, tau, to bind various targets such as 14-3-3, protein kinase C, synphilin-1, Elk and Tat-binding protein 1. The alpha and beta synucleins are found primarily in brain tissue where they are seen mainly in neuronal cytosol and presynaptic terminals. The antibody specific for alpha-synuclein does not cross react with beta-synuclein and will be useful for the confirmation of Parkinson's disease and Lewy body dementia and other studies for the localization and detection of alpha-synuclein. Four alternatively spliced transcripts that encode two different isoforms of the alpha-synuclein protein have been identified.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Bioinformatics

Protein Aliases: Alpha synuclein; Alpha-synuclein; MGC110988; MGC110988 antibody; NACP; NACP antibody; non A-beta component of AD amyloid; Non-A beta component of AD amyloid; Non-A4 component of amyloid precursor; OTTHUMP00000218549; OTTHUMP00000218551; OTTHUMP00000218552; OTTHUMP00000218553; OTTHUMP00000218554; PARK 1; PARK 1 antibody; PARK 4; PARK 4 antibody; PARK1 antibody; PARK4 antibody; PD 1 antibody; PD1 antibody; SNCA antibody; synuclein alpha-140; synuclein, alpha (non A4 component of amyloid precursor); SYUA

Gene Aliases: NACP; PARK1; PARK4; PD1; SNCA

UniProt ID: (Human) P37840, (Rat) P37377

Entrez Gene ID: (Human) 6622, (Rat) 29219

Molecular Function: membrane trafficking regulatory protein