Invitrogen™ Alpha-synuclein Monoclonal Antibody (LB509)

Catalog No.	Quantity
18-021-5	1 mL

Catalog No. 18-021-5 Supplier Invitrogen™ Supplier No. 180215

Description

Mouse Monoclonal Antibody

Synucleins (α, β, and γ) are a family of cytoplasmic proteins found predominantly, and abundantly, in the brain with cellular distribution concentrated in presynaptic nerve terminals, primarily near vesicles. Synucleins are thought to be involved in neuronal plasticity, synaptic function, and neurodegenerative disease. α-Synuclein (~18 kDa) is abundant in Lewy Bodies in sporadic Parkinson's Disease (PD) and Dementia with Lewy Bodies (DLB). It has been reported that the NAC fragment from α-synuclein is a major component of amyloid plaques in Alzheimer's Disease. LB509 reacts with α-synuclein but not ^SM-synuclein. Reactivity has not been observed with γ-synuclein. Positive tissue: Parkinson's disease tissue. Expected Staining Pattern: Cytoplasmic.

Alpha-synuclein is a member of the synuclein family, which also includes beta- and gamma-synuclein. These proteins are abundant in the brain and play a role in regulating synaptic transmission and membrane trafficking. Alpha- and beta-synuclein have been shown to selectively inhibit phospholipase D2, suggesting that they may also be involved in signaling pathways. Defects in the SNCA gene, which encodes alpha-synuclein, have been implicated in the pathogenesis of Parkinson's disease. Additionally, alpha-synuclein peptides are a major component of amyloid plaques in the brains of patients with Alzheimer's disease. Multiple alternatively spliced transcripts encoding different isoforms have been identified for this gene. Diseases associated with SNCA include Parkinson Disease 1, Autosomal Dominant and Dementia, Lewy Body.

Specifications

• Antigen: Alpha-synuclein
• Applications: Immunohistochemistry (Paraffin)
• Classification: Monoclonal
• Clone: LB509
• Conjugate: Unconjugated
• Dilution: Concentrate Antibody
• Formulation: PBS with 1% BSA and 0.1% sodium azide
• Gene: SNCA
• Gene Accession No.: P37840
• Gene Alias: Alpha synuclein; alphaSYN; alpha-synuclein; NACP; non A-beta component of AD amyloid; non-A beta component of AD amyloid; non-A4 component of amyloid; Non-A4 component of amyloid precursor; PARK1; PARK4; PD1; Snca; Syn; synuclein alpha; synuclein alpha-140; synuclein, alpha; synuclein, alpha (non A4 component of amyloid precursor)
• Gene Symbols: SNCA
• Host Species: Mouse
• Immunogen: Lewy Bodies purified from patients suffering dementia with Lewy Bodies.
• Purification Method: Purified
• Quantity: 1 mL
• Regulatory Status: RUO
• Primary or Secondary: Primary
• Gene ID (Entrez): 6622
• Target Species: Human
• Content And Storage: 2-8°C
• Product Type: Antibody
• Form: Liquid
• Isotype: IgG1 κ
• Host abrv: Ms
• Reactivity: Human
• Shipping Condition: Wet Ice
• Reactivity abrv: Hu
• Target Gene: alpha-Synuclein
• Validated Application abrv: IHC (FFPE)

Frequently Asked Questions (FAQs)

I know that there's no single procedure that applies to all antigens, but what are some commonly used procedures for performing immunohistochemical staining of brain tissue?

Here is a procedure for the immunohistochemical staining of Beta-amyloid with paraffin embedded sections of transgenic mouse brain. The following protocol was developed for use with paraffin embedded sections stained with a variety of our antibodies against Beta-amyloid. Applicable rabbit and mouse anti-Beta-amyloid antibodies are: 51-2700, 13-0200, 71-5800, 13-0100Z, 37-4200, 43-7900, 44-136, 700254, 36-6900, and AHB0121.

Beta-amyloid Staining:
- Transgenic mice (expressing the transgenes PS1-A246E + APP swe, APP swe alone, or PS1-A246E alone) were perfused with 1 x Dulbecco's phosphate buffered saline (D-PBS) followed by 4% paraformldehyde buffered with D-PBS. Brain tissues were then embedded in paraffin prior to microtome sectioning.
- After mounting on slides, the paraffin-embedded tissue sections were then deparaffinized with heat. The sections were then incubated for 3 minutes in 70% formic acid. Next, they were deparaffinized further with xylene followed with 100% ethanol. The sections were then re-hydrated in a graded ethanol series (100% ethanol, 95% ethanol, 70% ethanol, and then water).
- Endogenous peroxidase activity was quenched with 3% hydrogen peroxide in methanol. The sections were then heated in the microwave for 5-7 minutes in water, cooled at room temperature for 5 minutes and then rinsed in water. The sections were then washed in TBS (0.05 M Tris-HCl, pH 7.6, with 0.25 M NaCl) prior to blocking. Non-specific binding was blocked with 3% normal goat serum and 0.1% Triton X-100 in TBS for 1 hour at room temperature.
- The sections were then stained with anti-Beta-amyloid antibodies. The staining solution typically consisted of the antibody at a concentration of 5 µg/mL in TBS containing 2% normal goat serum. After incubation at room temperature for at least 1 hr., the sections were washed in TBS 3 times for 5 minutes each. An anti-mouse or anti-rabbit secondary antibody labeled with HRP was then used with DAB or AEC as substrates to stain the Beta-amyloid.
[Adapted from Borchelt, D.R., et al. (1997) Accelerated amyloid deposition in the brains of transgenic mice coexpressing mutant presenilin 1 and amyloid precursor proteins. Neuron 19:939-945.]

Tau and Synuclein Staining:
Here is a general procedure for the immunohistochemical staining of tau, its phosphorylated forms, and synuclein in frozen sections of rat brain. This protocol is adapted from one kindly contributed by Dr. Emil Adamec, M.D., Ph.D., McLean Hospital, Belmont, MA. The procedure was developed for use with cryostat sections of rat brain.
- Sections were fixed with 4% paraformaldehyde for 25 minutes. Sections were then extracted with 0.01% (v/v) Triton X-100. Endogenous peroxidase was blocked with 3% hydrogen peroxide. Sections were then treated with 80% formic acid for 5 to 10 minutes at room temperature to enhance staining. Note that formic acid treatment is also very useful prior to staining with anti-synuclein antibodies.
- The various antibodies were used at dilutions of 1:100 to 1:250. The sections were incubated with the diluted primary antibody overnight at 4°C. A species-specific secondary antibody labeled with HRP was then used with either DAB or AEC to stain the antigens.
- Applicable rabbit and mouse anti-tau and phospho-tau antibodies are: AHB0042, AHB0061, 44-738G, 39-1800, 13-6400, 13-1400, and 18-7461. Anti-synuclein antibodies useful for IHC are: 18-0215, 18-7461, 32-8100, 32-8200, 32-8500, 35-8300, 35-8400, 39-1800, and AHB0261.