Overview

  • Product name

    Human Alpha-synuclein ELISA Kit
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Serum 5 1.8%
    Inter-assay
    Sample n Mean SD CV%
    Serum 3 2.7%
  • Sample type

    Cell culture supernatant, Serum, Cell culture extracts, Tissue Extracts, Heparin Plasma, EDTA Plasma, Citrate Plasma, Cerebral Spinal Fluid
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    110 pg/ml
  • Range

    281 pg/ml - 3200 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell culture supernatant 104 97% - 116%
    Serum 100 94% - 103%
    Cell culture extracts 105 104% - 107%
    Tissue Extracts 107 104% - 112%
    Heparin Plasma 103 93% - 114%
    EDTA Plasma 95 67% - 115%
    Citrate Plasma 98 94% - 105%
    Cerebral Spinal Fluid 118 107% - 132%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Human
  • Product overview

    Alpha-synuclein in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Alpha-synuclein protein in human serum, plasmas, cerebrospinal fluid, cell culture supernatants, and cell and tissue extracts. (UniprotID: P37840)


    The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material.  TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.


    This kit is knockout validated for specifically detecting alpha-synuclein in human HAP1 cells.

  • Notes

    Alpha-synuclein is a 140 amino acid protein encoded by the SNCA gene, primarily found in neural tissue. It is a member of the synuclein family of proteins including β-Synuclein and γ-Synuclein. Alpha-synuclein has been found concentrated in the presynaptic nerve terminals of neurons, and has been found in the nucleus of neurons. Several physiological roles for Alpha-synuclein have been suggested, and is a major component of Lewy Bodies. This protein appears to play a role in dopamine metabolism and vesicle trafficking.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Pre-coated microplate (12 x 8 well strips)

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Human Alpha Synuclein Capture Antibody 1 x 600µl
    10X Human Alpha Synuclein Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml
    5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml
    Antibody Diluent CPI 1 x 6ml
    Human Alpha Synuclein Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS 1 x 50ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • Research areas

  • Function

    May be involved in the regulation of dopamine release and transport. Induces fibrillization of microtubule-associated protein tau. Reduces neuronal responsiveness to various apoptotic stimuli, leading to a decreased caspase-3 activation.
  • Tissue specificity

    Expressed principally in brain but is also expressed in low concentrations in all tissues examined except in liver. Concentrated in presynaptic nerve terminals.
  • Involvement in disease

    Genetic alterations of SNCA resulting in aberrant polymerization into fibrils, are associated with several neurodegenerative diseases (synucleinopathies). SNCA fibrillar aggregates represent the major non A-beta component of Alzheimer disease amyloid plaque, and a major component of Lewy body inclusions. They are also found within Lewy body (LB)-like intraneuronal inclusions, glial inclusions and axonal spheroids in neurodegeneration with brain iron accumulation type 1.
    Parkinson disease 1
    Parkinson disease 4
    Dementia Lewy body
  • Sequence similarities

    Belongs to the synuclein family.
  • Domain

    The 'non A-beta component of Alzheimer disease amyloid plaque' domain (NAC domain) is involved in fibrils formation. The middle hydrophobic region forms the core of the filaments. The C-terminus may regulate aggregation and determine the diameter of the filaments.
  • Post-translational
    modifications

    Phosphorylated, predominantly on serine residues. Phosphorylation by CK1 appears to occur on residues distinct from the residue phosphorylated by other kinases. Phosphorylation of Ser-129 is selective and extensive in synucleinopathy lesions. In vitro, phosphorylation at Ser-129 promoted insoluble fibril formation. Phosphorylated on Tyr-125 by a PTK2B-dependent pathway upon osmotic stress.
    Hallmark lesions of neurodegenerative synucleinopathies contain alpha-synuclein that is modified by nitration of tyrosine residues and possibly by dityrosine cross-linking to generated stable oligomers.
    Ubiquitinated. The predominant conjugate is the diubiquitinated form.
    Acetylation at Met-1 seems to be important for proper folding and native oligomeric structure.
  • Cellular localization

    Cytoplasm, cytosol. Membrane. Nucleus. Cell junction, synapse. Secreted. Membrane-bound in dopaminergic neurons.
  • Information by UniProt
  • Alternative names

    • Alpha synuclein
    • Alpha-synuclein
    • Alpha-synuclein, isoform NACP140
    • alphaSYN
    • MGC105443
    • MGC110988
    • MGC127560
    • MGC64356
    • NACP
    • Non A beta component of AD amyloid
    • Non A4 component of amyloid
    • Non A4 component of amyloid precursor
    • Non-A beta component of AD amyloid
    • Non-A-beta component of alzheimers disease amyloid , precursor of
    • Non-A4 component of amyloid precursor
    • Non-A4 component of amyloid, precursor of
    • OTTHUMP00000218549
    • OTTHUMP00000218551
    • OTTHUMP00000218552
    • OTTHUMP00000218553
    • OTTHUMP00000218554
    • PARK 1
    • PARK 4
    • PARK1
    • PARK4
    • Parkinson disease (autosomal dominant, Lewy body) 4
    • Parkinson disease familial 1
    • SNCA
    • Snca synuclein
    • Snca synuclein, alpha (non A4 component of amyloid precursor)
    • SYN
    • Synuclein alpha
    • Synuclein alpha 140
    • Synuclein, alpha (non A4 component of amyloid precursor)
    • SYUA_HUMAN
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab210973 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Sandwich ELISA Use at an assay dependent concentration.

Images

  • SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

     

  • Background-subtracted data values (mean +/- SD) are graphed.

  • Background-subtracted data values (mean +/- SD) are graphed.

  • The concentrations of Alpha Synuclein were measured in duplicates, interpolated from the Alpha Synuclein standard curves and corrected for sample dilution. Undiluted samples are as follows:  serum 67%, plasma (citrate) 30%, plasma (EDTA) 30%, and plasma (heparin) 30%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).  The mean Alpha Synuclein concentration was determined to be 2.7 ng/mL in serum, 9.0 ng/mL in plasma (citrate), 9.2 ng/mL in plasma (EDTA), and 7.0 ng/mL in plasma (heparin).

  • The concentrations of Alpha Synuclein were measured in duplicates, interpolated from the Alpha Synuclein standard curves and corrected for sample dilution. Undiluted samples are as follows:  cerebrospinal fluid 33%, culture media 50%.  The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). 

  • The concentrations of Alpha Synuclein were measured in duplicate and interpolated from the Alpha Synuclein standard curve and corrected for sample dilution.  The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).  The mean Alpha Synuclein concentration was determined to be 16 ng/mg in SH-SY5Y cell extract, 83 µg/mg in human brain extract, and 115 µg/mg in human fetal brain extract.

  • Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).  The mean Alpha Synuclein concentration was determined to be 2.45 ng/mL with a range of 1.32 – 5.53 ng/mL.

  • Raw data is plotted for wild-type and alpha synuclein-knockout HAP1 cell extracts tested at 111 μg/mL.

Protocols

References

This product has been referenced in:

  • Zou W  et al. Blocking meningeal lymphatic drainage aggravates Parkinson's disease-like pathology in mice overexpressing mutated a-synuclein. Transl Neurodegener 8:7 (2019). Read more (PubMed: 30867902) »
  • Svarcbahs R  et al. Removal of prolyl oligopeptidase reduces alpha-synuclein toxicity in cells and in vivo. Sci Rep 8:1552 (2018). Read more (PubMed: 29367610) »
See all 2 Publications for this product

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