SNAP25 Antibody (JE45-86)

Western Blot: SNAP25 Antibody (JE45-86) [NBP3-32961] - Western blot analysis of SNAP25 on different lysates with Rabbit anti-SNAP25 antibody (NBP3-32961) at 1/2,000 dilution. Lane 1: SH-SY5Y cell lysate (20 ug/Lane) Lane 2: Neuro-2a cell lysate (20 ug/Lane) Lane 3: K-562 cell lysate (negative) (20 ug/Lane) Lane 4: Human brain tissue lysate (40 ug/Lane) Predicted band size: 23 kDa Observed band size: 25 kDa Exposure time: 1 minute 21 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 (NBP3-32961) at 1/2,000 dilution was used in 5% NFDM/TBST at 4C overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.

Immunohistochemistry: SNAP25 Antibody (JE45-86) [NBP3-32961] - Immunohistochemical analysis of paraffin-embedded human brain tissue with Rabbit anti-SNAP25 antibody (NBP3-32961) at 1/500 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 (NBP3-32961) at 1/500 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.

Immunocytochemistry/ Immunofluorescence: SNAP25 Antibody (JE45-86) [NBP3-32961] - Immunocytochemistry analysis of SH-SY5Y cells labeling SNAP25 with Rabbit anti-SNAP25 antibody (NBP3-32961) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-SNAP25 antibody (NBP3-32961) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.

Flow Cytometry: SNAP25 Antibody (JE45-86) [NBP3-32961] - Flow cytometric analysis of SH-SY5Y cells labeling SNAP25. Cells were fixed and permeabilized. Then stained with the primary antibody (NBP3-32961, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

• Reactivity: Hu, Mu, Rt
• Applications: WB, Flow, ICC/IF, IHC
• Clone: JE45-86
• Clonality: Monoclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: 1 mg/ml

SNAP25 Antibody (JE45-86) Summary

• Description: Novus Biologicals Rabbit SNAP25 Antibody (JE45-86) (NBP3-32961) is a recombinant monoclonal antibody validated for use in IHC, WB, Flow and ICC/IF. All Novus Biologicals antibodies are covered by our 100% guarantee.
• Additional Information: Recombinant Monoclonal Antibody
• Immunogen: Recombinant protein within Human SNAP25 aa 51-150 / 206. (Uniprot: P60880)
• Localization: Cytoplasm, perinuclear region. Cell membrane.
• Source: HEK293
• Isotype: IgG
• Clonality: Monoclonal
• Host: Rabbit
• Gene: SNAP25
• Purity: Protein A purified

Applications/Dilutions

• Dilutions:
  • Flow Cytometry 1:1000
  • Immunocytochemistry/ Immunofluorescence 1:100
  • Immunohistochemistry
  • Immunohistochemistry-Paraffin 1:500-1:5000
  • Western Blot 1:2000
• Theoretical MW: 23 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: 1*TBS (pH7.4), 0.05% BSA and 40% Glycerol
• Preservative: 0.05% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Protein A purified

Alternate Names for SNAP25 Antibody (JE45-86)

• bA416N4.2
• dJ1068F16.2
• FLJ23079
• resistance to inhibitors of cholinesterase 4 homolog
• RIC4
• RIC-4
• SEC9
• SNAP25
• SNAP-25SNAP
• SUP
• Super protein
• Synaptosomal-associated 25 kDa protein
• synaptosomal-associated protein 25
• synaptosomal-associated protein, 25kD
• synaptosomal-associated protein, 25kDa

Background

SNAP25 (Synaptosomal associated protein of 25 kDa) is a presynaptic plasma membrane protein that is widely distributed throughout the brain and involved in the regulation of neurotransmitter release. Decreased levels of SNAP25 have been found in the brains of patients with Down Syndrome and Alzheimers Disease (Greber et al.,1999). In addition, a significant reduction in the hippocampal expression of SNAP25 has also been found in patients with Schizophrenia (Fatemi et al., 2001).

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Bioinformatics

• Gene Symbol: SNAP25