Anti α synuclein

Manufactured by Abcam

Sourced in United Kingdom, United States

Anti-α-synuclein is a lab equipment product that can be used to detect and study the α-synuclein protein. α-synuclein is a small protein found in the presynaptic terminals of neurons and is associated with neurodegenerative diseases such as Parkinson's disease.

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Lab products found in correlation

Close spelling variants of this product

α-synuclein (15 citations) Anti-α-synuclein antibody (5 citations) Anti-pS129-α-synuclein (3 citations) Anti-p-α-synuclein (3 citations)

12 protocols using anti α synuclein

Quantitative Immunoblot Analysis of Signaling Proteins

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Protein extracts from tumor tissues and cells were prepared in radioimmunoprecipitation assay (RIPA) buffer containing protease inhibitors (Sigma-Aldrich). Protein samples were separated by sodium dodecyl sulphate–polyacrylamide gel electrophoresis and transferred onto nitrocellulose membranes. Primary antibodies (1:500 dilution) used here are as follows: anti-Akt, anti-Akt (ser473), anti-mTOR, anti-phospho-mTOR (Ser2448), anti-phospho-p70S6K1 (Thr389), anti-p70S6K1, anti-phospho-4EBP (Thr37/46), anti-4EBP (Cell Signaling Technology, Beverly, MA, USA), anti-α-synuclein (Abcam, Cambridge, MA, USA), and anti-β-actin (Sigma-Aldrich). Blots were developed with an enhanced chemiluminescence system (GE Healthcare Biosciences, Piscataway, NJ, USA). Densitometric analysis of the blots were performed using the Quantity One software (Bio-Rad Laboratories, Richmond, CA, USA).

Ge Y, & Xu K. (2016). Alpha-synuclein contributes to malignant progression of human meningioma via the Akt/mTOR pathway. Cancer Cell International, 16, 86.

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Comprehensive Antibody Panel for Alzheimer's Research

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anti-Aβ/APP 4G8 (mouse mAb, Covance), anti-Aβ/APP 6E10 (mouse mAb, Covance), anti-AβOs scFvA13 and scFvIm3 (ref. 13 (link)), anti-oligomer A11 (ref. 21 (link)) (rabbit pAb Millipore), anti-Aβ (rabbit mAb, Cell Signaling), anti α-Synuclein (mouse mAb, Abcam), anti-APP C-terminal fragment (rabbit pAb, Sigma), anti-APP N-terminal 22C11 (mouse mAb, Millipore), anti-V5 (mouse mAb Invitrogen, and rabbit pAb Sigma), anti-V5-HRP (mouse mAb, Sigma), anti-His tag (mouse mAb Millipore), anti-KDEL (mouse mAb, Assay Design/Stressgen) (kind gift of Prof. R. Sitia, HSR San Raffaele, Milan), anti-mTOR (rabbit pAb, Cell Signaling), anti-phospho-mTOR (rabbit pAb, Cell Signaling), anti-P70S6K (rabbit pAb, Cell Signaling) (kind gift of Dr G. Amadoro, CNR, Rome), anti-phospho-P70S6K (rabbit pAb, Cell Signaling), anti ERK1/2 (rabbit pAb, Cell Signaling), anti phosho-ERK1/2 (rabbit pAb, Cell Signaling), anti phospho-CREB (rabbit pAb, Cell Signaling), anti β-Actin (mouse mAb and rabbit pAb, Sigma), anti-calnexin (rabbit pAb, Sigma), anti-GM130 (mouse mAb, Covance), anti-Golgi 58K (mouse mAb, Sigma), anti-Rab3A (mouse mAb, Sigma), anti-Rab5 (rabbit pAb, Abcam), anti-Rab7 (goat pAb, Santa Cruz), anti-TGN46 (rabbit, pAb), anti-LAMP-1 (rabbit pAb, Abcam). anti-Rab5, anti-Rab7, anti-TGN46 are kind gift of Dr V. Triaca, CNR, Rome.

Meli G., Lecci A., Manca A., Krako N., Albertini V., Benussi L., Ghidoni R, & Cattaneo A. (2014). Conformational targeting of intracellular Aβ oligomers demonstrates their pathological oligomerization inside the endoplasmic reticulum. Nature Communications, 5, 3867.

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Quantitative Protein Expression Analysis

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The bicin-choninic Acid (BCA) Protein Assay Kit (CW2011S, CWBIO, Beijing, China) was used to calculate protein concentration. The loading buffer was added to the protein supernatant, which was collected as described in the ELISA test protocol, heated for 5 min (95 °C), loaded on 10% acrylamide–SDS gel (P0056B, Shanghai, China), and transferred to a polyvinylidene fluoride (PVDF) membrane (Millipore, Billerica, MA, USA). The molecular weight of the protein was marked by a protein marker (26616, Thermo, Waltham, MA, USA). The membrane was blocked with 5% skim milk in TBST for 2 h at room temperature. Primary antibodies anti-α-synuclein (Abcam, ab155038, Cambridge, UK), anti-olig2 (CST, 17198, Boston, MA, USA), anti-Iba1 (CST, ab109186, Boston, MA, USA), and anti-β-actin (Bioss, bs-0061R, Beijing, China) were incubated overnight at 4 °C and washed. Species-appropriate, HRP-conjugated secondary antibodies were incubated for 1 h at room temperature and washed three times. Enhanced chemiluminescence reagents reacted with HRP, and the signal intensity value was read by the ultra-sensitive chemiluminescence system (Fusion FX.EDGE, VILBER Smart Imaging, Paris, France). The integrated density ratio of α-synuclein: β-actin represents the expression level of α-synuclein, as do the olig2 and Iba1.

Ma X., Chen L., Song N., Qu L., Wang J, & Xie J. (2021). Blood-Derived α-Synuclein Aggregated in the Substantia Nigra of Parabiotic Mice. Biomolecules, 11(9), 1287.

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Western blot and immunoprecipitation of brain proteins

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Dissected mouse brain tissues were lysed in RIPA buffer and the protein homogenates were then separated by SDS-PAGE and transferred onto a polyvinylidene fluoride membrane. After blocking, the membranes were incubated overnight with primary antibodies, followed by washing and incubation with horseradish peroxidase conjugated secondary antibody for 1 h. Immunoreactive bands were then visualized using the ECL reagent (WBKLS0100, Merck Millipore). The densities of the immunoblotting bands were semiquantified using Image J software (National Institutes of Health). The primary antibodies used were as follows: anti-tyrosine hydroxylase (TH) (#T2928, Sigma), anti-α-synuclein (#ab1903, Abcam), anti-SIRT1 (#07–131, Millipore), anti-p62 (#18420–1-AP, Protein Tech Group), anti-LC3B (#18725–1-AP, Protein Tech Group), anti-acetyl-Lysine (#05–515, Millipore), anti-caspase-3 (#9662, CST), and anti-GAPDH (#10494–1-AP, Protein Tech Group).
For immunoprecipitation, extracted proteins were incubated with anti-LC3B (#3868, CST) at 4°C overnight with rotation, followed by precipitation with protein A agarose beads (P2012, Beyotime). Immunoprecipitates were then washed and subjected to SDS-PAGE separation. Subsequent immunoblotting procedures were the same as described above.

Guo Y.J., Dong S.Y., Cui X.X., Feng Y., Liu T., Yin M., Kuo S.H., Tan E.K., Zhao W.J, & Wu Y.C. (2016). Resveratrol alleviates MPTP-induced motor impairments and pathological changes by autophagic degradation of α-synuclein via SIRT1-deacetylated LC3. Molecular nutrition & food research, 60(10), 2161-2175.

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Immunofluorescence Staining of Neuronal Markers

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Cells were seeded on matrigel-coated glass coverslips and they were fixed for 20 min in ice in 4% paraformaldehyde (PFA, Sigma), solution in phosphate-buffered saline (PBS, Euroclone). Then, cells were permeabilized for 30 min in a blocking solution, containing 0.5% Triton X-100 (Sigma-Aldrich) and 10% donkey serum (Sigma-Aldrich), and incubated overnight at 4 °C with the primary antibodies in a blocking solution. Then, cells were washed with PBS and incubated for 1 h at room temperature with Hoechst and with secondary antibodies. The following antibodies were used: anti-OCT4 (1:100, Abcam), anti-NANOG (1:100, Abcam), anti-FOXA2 (1:300, Abcam), anti-NESTIN (1:300 Millipore), anti-TH (1:200, Immunological Sciences), anti-MAP2 (1:500, Immunological Sciences), anti-TOMM20 (1:300, Novus), anti-α-Synuclein (clone LB509, 1:100, Thermo), anti-GFP (1:500, Thermo), anti-α-Synuclein (phosphoS129, 1:300, Abcam), anti-TAU (1:500, Millipore), anti-LAMP1 (1:500, Abcam), anti-Synapsin-1 (1:500, Synaptic Systems), anti-SMI311 (1:500, BioLegend), anti GM130 (1:300, BD), anti-GRIM19 (1:300, Abcam). All the secondary antibodies used for the immunofluorescence staining are Alexa Fluor^TM.

Iannielli A., Luoni M., Giannelli S.G., Ferese R., Ordazzo G., Fossati M., Raimondi A., Opazo F., Corti O., Prehn J.H., Gambardella S., Melki R, & Broccoli V. (2022). Modeling native and seeded Synuclein aggregation and related cellular dysfunctions in dopaminergic neurons derived by a new set of isogenic iPSC lines with SNCA multiplications. Cell Death & Disease, 13(10), 881.

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Molecular Mechanisms in Neurodegenerative Diseases

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For the experiments, we used the following materials: culture media (Gibco-Thermo Fisher Scientific, Waltham, MA, USA); fetal bovine serum and minimum essential medium with Earle’s salt (EuroClone Life Science, Pero, MI, Italy); penicillin/streptomycin and RIPA buffer (Sigma-Aldrich, St. Louis, MO, USA); a protease inhibitor cocktail (Roche, Basilea, Switzerland); Quick Start™ Bradford Dye Reagent and Clarity™ Western ECL Blotting Substrates; iScript^TM cDNA synthesis, retro-transcription kit (BioRad, Segrate, MI, Italy); ReliaPrep™ Miniprep RNA Extraction System and GoTaq qPCR Master Mix (Promega, Madison, WI, USA); SYBR Green (Takara, Kyoto, Japan); and synthetic oligonucleotides from Eurofins Genomics (Edersberg, Germany). The primary antibodies used were as follows: anti-Nrf2 (ElabScience, Houston, TX, USA), anti-β-Actin (Sigma-Aldrich, St. Louis, MO, USA), anti-Tfeb (ab2636, Abcam, Cambridge, UK), anti-α-synuclein (ab138501 Abcam, Cambridge, UK), and anti-LAMP2 (Santa Cruz Biotechnology, Dallas, TX, USA). The secondary antibodies were purchased from Jackson Laboratories (Bar Harbor, ME, USA). The secondary antibodies for immunofluorescence were as follows: Alexa Fluor 488 and Alexa Fluor 546; these were purchased from Thermo Fisher, and the Human MDA (Malondialdehyde) ELISA Kit was purchased from Fine Test, Labospace (Milan, MI, Italy).

Mingione A., Pivari F., Plotegher N., Dei Cas M., Zulueta A., Bocci T., Trinchera M., Albi E., Maglione V., Caretti A., Bubacco L., Paroni R., Bottai D., Ghidoni R, & Signorelli P. (2021). Inhibition of Ceramide Synthesis Reduces α-Synuclein Proteinopathy in a Cellular Model of Parkinson’s Disease. International Journal of Molecular Sciences, 22(12), 6469.

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Protein Expression Analysis in Mouse Brain

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After the mice were sacrificed at the indicated time point, the distal colon and the middle brain substantia nigra were rapidly removed and lysed in RIPA lysis buffer (50 mM Tris–HCl, pH 8.0; 1% NP-40; 0.5% sodium deoxycholate; 150 mM NaCl; 0.1% SDS) containing protease and phosphatase inhibitor cocktails and 1 mM phenyl-methylsulphonyl fluoride (PMSF). Equal amounts of protein (30 μg) were loaded per lane and separated by 10% SDS-PAGE. Proteins in the gels were transferred to immobilon polyvinylidene difluoride (PVDF) membrane. The membrane was blocked with 3% BSA and incubated with primary antibody overnight at 4°C (anti-α-synuclein, Abcam, 1:1,000; anti-TH, Santa Cruz, 1:500; anti-phospho-α-synuclein (phospho S129), Abcam 1:100; anti-MyD88, Cell Signaling, 1:1,000; anti-NF-κB, Santa Cruz, 1:1,000; anti-β-actin, Sigma, 1:5,000) followed by secondary antibodies conjugated to horseradish peroxidase (Santa Cruz Biotechnology, CA, USA) and visualized using chemiluminescent detection methods (Amersham Company).

Yang X., Qian Y., Xu S., Song Y, & Xiao Q. (2018). Longitudinal Analysis of Fecal Microbiome and Pathologic Processes in a Rotenone Induced Mice Model of Parkinson’s Disease. Frontiers in Aging Neuroscience, 9, 441.

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Striatum Protein Analysis by Western Blot

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Striatum was homogenized in RIPA buffer containing protease and phosphatase inhibitors (1 mM sodium orthovanadate, 1 mM aprotinin, and 1% protease inhibitor cocktail) and centrifuged at 10000g for 10 minutes at 4°C. Supernatants were used for western blotting, as described in da Rosa-Junior.^{33 (link)} The following primary antibodies were used: anti-phospho-JNK and anti-JNK were purchased from R&D Systems, Minnesota; anti-Bok, anti-Bcl-xL, anti-caspase-9, anti-caspase-3, anti-cleaved caspase-3, anti-phospho-p38, anti-p38, anti-phospho-ERK1/2, and anti-ERK1/2 from Cell Signaling, Massachusetts; anti-superoxide dismutase 1 (SOD1), anti-heme oxygenase-1 (HO-1), and anti-α-synuclein from Abcam, Cambridge, UK; anti-catalase (CAT) was from Merk Millipore, Massachusetts; anti-GSK-3β from Santa Cruz Biotechnology, Texas; anti-β-actin from Sigma-Aldrich, Missouri.

Glänzel N.M., Grings M., da Rosa-Junior N.T., Cereta de Carvalho L.M., Mohsen A.W., Wipf P., Wajner M., Vockley J, & Leipnitz G. (2020). The mitochondrial-targeted reactive species scavenger JP4-039 prevents sulfite-induced alterations in antioxidant defenses, energy transfer, and cell death signaling in striatum of rats. Journal of inherited metabolic disease, 44(2), 481-491.

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Hippocampal Protein Analysis via Western Blot

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Hippocampus blocks (~100 mg) were harvested, immediately soaked in liquid nitrogen, smashed into power, dissolved in SDS lysis buffer (50 mM Tris-HCl, pH7.0, 2% SDS, 6% glycerol, 100 mM DTT) containing a protease inhibitor mixture, and subjected to SDS-PAGE. The protein bands were transferred to an Immun-Blot PVDF membrane (Bio-Rad). Western blot analyses were performed as described previously (Baker et al., 2015 (link)). Antibodies used for western blot including anti-α-synuclein (1:1000) and anti-β-amyloid (1:1000) were purchased from Abcam. The anti-Parkin (Prk8) (1:1000) and anti-SQSTM1/p62 (1:1000) antibodies were purchased from Cell Signaling Technology. The anti-β-actin (1:2000) antibody was purchased from Santa Cruz Biotechnology.

Chen K., Yuan R., Geng S., Zhang Y., Ran T., Kowalski E., Liu J, & Li L. (2016). Toll-interacting protein deficiency promotes neurodegeneration via impeding autophagy completion in high-fat diet-fed ApoE−/− mouse model. Brain, behavior, and immunity, 59, 200-210.

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Monoclonal and Polyclonal Antibodies for Research

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The following monoclonal antibodies were purchased: anti-HSP70, anti-p-ERK, and anti-α-synuclein (Abcam); anti-GAPDH (Santa Cruz Biotechnology); anti-VHR (BD Biosciences); anti-HA (Roche); anti-Aβ (BioLegend); anti-NeuN (Merck Millipore); anti-HSP70, anti-GST, anti-cleaved caspase-3, anti-Bak, and anti-Bax (Cell Signaling Technology). The following polyclonal antibodies were also obtained: anti-PSD95 (Abcam); anti-HSP70, anti-VRK3, anti-VHR, anti-p-ERK, anti-ERK, and anti-His (Cell Signaling Technology); anti-VRK3 (Atlas antibodies); anti-HA (YPYDVPDYA; Bethyl Laboratories); anti-FLAG (D-8), anti-Lamin B, and anti-MAP2 (I-18) (Santa Cruz Biotechnology).

Song H., Kim W., Kim S.H, & Kim K.T. (2016). VRK3-mediated nuclear localization of HSP70 prevents glutamate excitotoxicity-induced apoptosis and Aβ accumulation via enhancement of ERK phosphatase VHR activity. Scientific Reports, 6, 38452.

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