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alpha+beta Synuclein Recombinant Rabbit Monoclonal Antibody [ST05-21]

Usd: 385

Specification

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Catalog# ET1609-66

alpha+beta Synuclein Recombinant Rabbit Monoclonal Antibody [ST05-21]

Application

  • WB

  • IF-Cell

  • IF-Tissue

  • IHC-P

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

Overview

Product Name

alpha+beta Synuclein Recombinant Rabbit Monoclonal Antibody [ST05-21]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human alpha+beta Synuclein aa 2-50 / 140.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IF-Tissue, IHC-P, FC

Target Molecular Weight

Predicted band size: 14 kDa

Positive Control

Mouse brain tissue lysate, rat brain tissue lysate, Hela, N2A, SH-SY5Y, human brain tissue, mouse brain tissue, rat brain tissue.

Conjugation

unconjugated

Clone Number

ST05-21

RRID

AB_3069875

Product Features

Form

Liquid

Concentration

1 mg/mL.(The concentration of this product may be batch-dependent)

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000-1:2,000

  • IF-Cell

  • 1:50-1:200

  • IF-Tissue

  • 1:50-1:200

  • IHC-P

  • 1:5,000

  • FC

  • 1:50-1:100

Target

Function

The synucleins, including α-synuclein (also designated NACP for nonamyloid component precursor), β-synuclein (also designated PNP 14 for phospho-neuroprotein 14) and γ-synuclein (also designated persyn or BCSG1 for breast cancer-specific gene 1) are presynaptic proteins abundant in neurons. Synucleins are predominantly expressed in the brain and are speculated to be involved in synaptic regulation and neuronal plasticity. α-Synuclein, identified as a component of Alzheimer’s disease amyloid plaques, is localized to neuronal cell bodies and synapses. Coordinate expression of α-synuclein and β-synuclein may be important during hematopoetic cell differentiation. A mutant form of α-synuclein is found in patients with early onset Parkinson’s disease. γ-Synuclein is associated with axonal pathology in Parkinson’s disease.

Background References

1. Hall H et al. Characterization of cognitive deficits in rats overexpressing human alpha-synuclein in the ventral tegmental area and medial septum using recombinant adeno-associated viral vectors. PLoS One 8:e64844 (2013).

2. Ejlerskov P et al. Tubulin polymerization-promoting protein (TPPP/p25a) promotes unconventional secretion of a-synuclein through exophagy by impairing autophagosome-lysosome fusion. J Biol Chem 288:17313-35 (2013).

Sequence Similarity

Belongs to the synuclein family.

Tissue Specificity

Highly expressed in presynaptic terminals in the central nervous system. Expressed principally in brain.

Post-translational Modification

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 (By similarity).; Acetylation at Met-1 seems to be important for proper folding and native oligomeric structure.

Subcellular Location

Cytoplasm, Nucleus, Membrane, Secreted, Cell junction.

UNIPROT

SwissProt: P37840 Human

SwissProt: Q16143 Human

SwissProt: O55042 Mouse

SwissProt: Q91ZZ3 Mouse

SwissProt: P37377 Rat

SwissProt: Q63754 Rat

Synonyms

Alpha synuclein antibody

Beta synuclein antibody

NACP antibody

Non A beta component of AD amyloid antibody

PARK1 antibody

PARK4 antibody

PD1 antibody

SNCA antibody

SNCB antibody

Synuclein alpha antibody

Expand

Images

  • Western blot analysis of alpha+beta Synuclein on different lysates with Rabbit anti-alpha+beta Synuclein antibody (<a href="/products/ET1609-66" style="font-weight: bold;text-decoration: underline;">ET1609-66</a>) at 1/1,000 dilution.<br /><br />Lane 1: Mouse brain tissue lysate<br />Lane 2: Rat brain tissue lysate<br /><br />Lysates/proteins at 40 µg/Lane.<br /><br />Predicted band size: 14 kDa<br />Observed band size: 15 kDa<br /><br />Exposure time: 43 seconds;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1609-66" style="font-weight: bold;text-decoration: underline;">ET1609-66</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of alpha+beta Synuclein on different lysates with Rabbit anti-alpha+beta Synuclein antibody (ET1609-66) at 1/1,000 dilution.

    Lane 1: Mouse brain tissue lysate
    Lane 2: Rat brain tissue lysate

    Lysates/proteins at 40 µg/Lane.

    Predicted band size: 14 kDa
    Observed band size: 15 kDa

    Exposure time: 43 seconds;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1609-66) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • ICC staining of alpha+beta Synuclein in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (<a href="/products/ET1609-66" style="font-weight: bold;text-decoration: underline;">ET1609-66</a>, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor&reg;488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

    ICC staining of alpha+beta Synuclein in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1609-66, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

  • ICC staining of alpha+beta Synuclein in N2A cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (<a href="/products/ET1609-66" style="font-weight: bold;text-decoration: underline;">ET1609-66</a>, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor&reg;488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

    ICC staining of alpha+beta Synuclein in N2A cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1609-66, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

  • ICC staining of alpha+beta Synuclein in SH-SY5Y cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (<a href="/products/ET1609-66" style="font-weight: bold;text-decoration: underline;">ET1609-66</a>, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor&reg;488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

    ICC staining of alpha+beta Synuclein in SH-SY5Y cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1609-66, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

  • Immunohistochemical analysis of paraffin-embedded human brain tissue with Rabbit anti-alpha+beta Synuclein antibody (<a href="/products/ET1609-66" style="font-weight: bold;text-decoration: underline;">ET1609-66</a>) at 1/5,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1609-66" style="font-weight: bold;text-decoration: underline;">ET1609-66</a>) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human brain tissue with Rabbit anti-alpha+beta Synuclein antibody (ET1609-66) at 1/5,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1609-66) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-alpha+beta Synuclein antibody (<a href="/products/ET1609-66" style="font-weight: bold;text-decoration: underline;">ET1609-66</a>) at 1/5,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1609-66" style="font-weight: bold;text-decoration: underline;">ET1609-66</a>) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-alpha+beta Synuclein antibody (ET1609-66) at 1/5,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1609-66) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-alpha+beta Synuclein antibody (<a href="/products/ET1609-66" style="font-weight: bold;text-decoration: underline;">ET1609-66</a>) at 1/5,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1609-66" style="font-weight: bold;text-decoration: underline;">ET1609-66</a>) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-alpha+beta Synuclein antibody (ET1609-66) at 1/5,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1609-66) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Flow cytometric analysis of alpha+beta Synuclein was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (<a href="/products/ET1609-66" style="font-weight: bold;text-decoration: underline;">ET1609-66</a>, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of alpha+beta Synuclein was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1609-66, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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