Human/Mouse/Rat SNAP25 Antibody

(1 citations)   
  • Species Reactivity
    Human, Mouse, Rat
  • Specificity
    Detects human, mouse, and rat SNAP25 in direct ELISAs and Western blots. In direct ELISAs, approximately 5% cross‑reactivity with recombinant human SNAP23 is observed.
  • Source
    Polyclonal Sheep IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant human SNAP25
    Asp140-Gly206
    Accession # P60880
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    1 µg/mL
    See below
  • Immunocytochemistry
    5-15 µg/mL
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Detection of Human and Rat SNAP25 by Western Blot. Western blot shows lysates of A172 human glioblastoma cell line, H4 human neuroglioma cell line, and PC‑12 rat adrenal pheochromocytoma cell line. PVDF Membrane was probed with 1 µg/mL of Sheep Anti-Human/Mouse/Rat SNAP25 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5946) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for SNAP25 at approximately 29 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.
Immunocytochemistry
SNAP25 in SH-SY5Y Human Cell Line. SH-SY5Y human neuroblastoma cells were cultured overnight in the presence of 1 mM Retinoic Acid (Catalog # 0695/50) prior to immersion fixation. SNAP25 was detected using a Sheep Anti-Human/Mouse/Rat SNAP25 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5946). The cells were stained with the NorthernLights 557-conjugated Donkey Anti-Sheep Affinity-purified IgG Secondary Antibody (red; Catalog # NL010). Actin filaments were stained with FITC-conjugated Phalloidin (green) and cell nuclei were counterstained with DAPI (blue). SNAP25 immunoreactivity was localized to the synaptic termini. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Immunocytochemistry
SNAP25 in PC-12 Rat Adrenal Pheochromocytoma Cells. PC-12 rat adrenal pheochromocytoma cells were cultured overnight in the presence of 50 ng/mL of Recombinant Rat b-NGF (Catalog # 556-NG) prior to immersion fixation. SNAP25 was detected using a Sheep Anti-Human/Mouse/Rat SNAP25 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5946). The cells were stained with the NorthernLights 557-conjugated Donkey Anti-Sheep Affinity-purified IgG Secondary Antibody (red; Catalog # NL010). The cell nuclei were counterstained with DAPI (blue). SNAP25 immunoreactivity was localized to the synaptic termini. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & Storage
    Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
    • 12 months from date of receipt, -20 to -70 °C as supplied.
    • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
    • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: SNAP25
SNAP25 (Synaptosomal associated protein of 25 kDa; also SUP) is a cytosolic 24-29 kDa member of the SNAP25 family of proteins. It is expressed in neurons, beta ‑cells, and adrenal chromaffin cells. SNAP25 is involved in vesicle exocytosis, where two presynaptic SNAP25, and one syntaxin‑1 molecule associates with vesicle membrane VAMP2 to form a alpha ‑helix via a zipper‑like mechanism. This approximates two membranes, which subsequently fuse to create a pore. Human SNAP25b is 206 amino acids (aa) in length. It contains two t‑SNARE coiled‑coil homology domains (aa 19‑81 and 140‑202) and a proteolytic cleavage site at Arg180:Ile181. Palmitoylation occurs between aa 85‑92; phosphorylation occurs on Thr138. There are three isoform variants. One shows nine aa changes over aa 58‑89 (isoform a), a second shows a 30 aa substitution for aa 1‑94, while a third contains an alternative start site at Met64. Full‑length human and mouse SNAP25 are identical in amino acid sequence.
  • Long Name:
    Synaptosomal-associated Protein, 25 kDa
  • Entrez Gene IDs:
    6616 (Human); 20614 (Mouse); 25012 (Rat)
  • Alternate Names:
    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
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

1 Citations: Showing 1 - 1

  1. SNAP23 Regulates Endothelial Exocytosis of von Willebrand Factor.
    Authors: Zhu Q, Yamakuchi M, Lowenstein C
    PLoS ONE, 2015;10(8):e0118737.
    Species: Human
    Sample Type: Whole Cells
    Application: IHC Not Specified
Cell and Tissue Staining Kits
Description Application Cat# Citations Images  

Anti-Sheep HRP-DAB Cell & Tissue Staining Kit

CTS019 1
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Isotype Controls
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Normal Sheep IgG Control

Ctrl 5-001-A 9
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Secondary Antibodies
Description Application Cat# Citations Images  

Sheep IgG HRP-conjugated Antibody

WB, Simple Western HAF016 14  
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Donkey Anti-Sheep IgG NorthernLights™ NL557-conjugated Antibody

Flow, IHC, ICC NL010 3
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Donkey Anti-Sheep IgG NorthernLights™ NL637-conjugated Antibody

Flow, IHC, ICC NL011 5
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Donkey Anti-Sheep IgG NorthernLights™ NL493-conjugated Antibody

Flow, IHC, ICC NL012 2
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Sheep IgG (H+L) PE-conjugated Antibody

Flow F0126  
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Donkey Anti-Sheep IgG Biotinylated Antibody

WB BAF016 2
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Sheep IgG (H+L) APC-conjugated Antibody

Flow F0127  
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Sheep IgG VisUCyte HRP Polymer Antibody

IHC VC006  
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Donkey Anti-Sheep IgG (H+L) Fluorescein-conjugated Antibody

Flow F0125 1
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Donkey Anti-Sheep IgG (H+L) PerCP-conjugated Antibody

Flow F0128
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