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Human/Mouse/Rat SNAP25 Antibody

Catalog #: AF5946
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AF5946
AF5946-SP
Detection of Human and Rat SNAP25 by Western Blot.
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Citations (1)
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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human/Mouse/Rat SNAP25 Antibody Summary

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 either lyophilized or as a 0.2 µm filtered solution in PBS.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
1 µg/mL
See below
Simple Western
10 µg/mL
Human/Mouse/Rat cerebellum
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.

Scientific Data

Western Blot Detection of Human and Rat SNAP25 antibody by Western Blot. View Larger

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 (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 antibody in SH-SY5Y Human Cell Line by Immunocytochemistry (ICC). View Larger

SNAP25 in SH-SY5Y Human Cell Line. SH-SY5Y human neuroblastoma cells were cultured overnight in the presence of 1 mM Retinoic Acid (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; 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 antibody in PC-12 Rat Adrenal Pheochromocytoma Cells by Immunocytochemistry (ICC). View Larger

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 (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; 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.

Simple Western View Larger

Detection of Human/Mouse/Rat SNAP25 by Simple WesternTM. Simple Western lane view shows lysates of human, mouse, rat cerebellum, loaded at 0.2 mg/mL. A specific band was detected for SNAP25 at approximately 31 kDa (as indicated) using 10 µg/mL of Sheep Anti-Human/Mouse/Rat SNAP25 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5946) followed by 1:50 dilution of HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). This experiment was conducted under reducing conditions and using the 2-40 kDa separation system.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
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

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Citation for Human/Mouse/Rat SNAP25 Antibody

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 Citation: Showing 1 - 1

  1. SNAP23 Regulates Endothelial Exocytosis of von Willebrand Factor.
    Authors: Zhu Q, Yamakuchi M, Lowenstein C
    PLoS ONE, 2015-08-12;10(8):e0118737.
    Species: Human
    Sample Types: Whole Cells
    Applications: IHC

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