Key Product Details

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Human

Applications

Validated:

Western Blot, Immunocytochemistry, Simple Western

Cited:

Immunohistochemistry

Label

Unconjugated

Antibody Source

Polyclonal Sheep IgG
View all SNAP25 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human SNAP25
Asp140-Gly206
Accession # P60880

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.

Clonality

Polyclonal

Host

Sheep

Isotype

IgG

Scientific Data Images for Human/Mouse/Rat SNAP25 Antibody

Detection of Human and Rat SNAP25 antibody by Western Blot.

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.

SNAP25 antibody in SH-SY5Y Human Cell Line by Immunocytochemistry (ICC).

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.

SNAP25 antibody in PC-12 Rat Adrenal Pheochromocytoma Cells by Immunocytochemistry (ICC).

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.

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.

Applications for Human/Mouse/Rat SNAP25 Antibody

Application
Recommended Usage

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed SH-SY5Y human neuroblastoma cell line and immersion fixed PC-12 rat adrenal pheochromocytoma cells

Simple Western

10 µg/mL
Sample: Human/Mouse/Rat cerebellum

Western Blot

1 µg/mL
Sample: A172 human glioblastoma cell line, H4 human neuroglioma cell line, and PC‑12 rat adrenal pheochromocytoma cell line

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

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.

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.

Calculators

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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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

Alternate Names

RIC4, SEC9, SUP

Entrez Gene IDs

6616 (Human); 20614 (Mouse); 25012 (Rat)

Gene Symbol

SNAP25

UniProt

P60880 (Human)

Additional SNAP25 Products

Product Documents for Human/Mouse/Rat SNAP25 Antibody

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human/Mouse/Rat SNAP25 Antibody

For research use only

Related Research Areas

Neuronal Markers

Citations for Human/Mouse/Rat SNAP25 Antibody

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Protocols

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