SNAP25 Recombinant antibody

SNAP25 Uni-rAb^TM Recombinant Antibody for IHC, IF/ICC, FC (Intra), ELISA

Host / Isotype

Rabbit / IgG

Reactivity

rat

Applications

IHC, IF/ICC, FC (Intra), ELISA

Conjugate

Unconjugated

CloneNo.

240069A1

Cat no : 83259-1-RR

Synonyms

SNAP 25, SNAP, RIC-4, RIC4, RIC 4

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Validation Data Gallery

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  IHC staining of mouse brain using 83259-1-RR

  Immunohistochemical analysis of paraffin-embedded mouse brain tissue slide using 83259-1-RR (SNAP25 antibody) at dilution of 1:200 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of mouse brain using 83259-1-RR

  Immunohistochemical analysis of paraffin-embedded mouse brain tissue slide using 83259-1-RR (SNAP25 antibody) at dilution of 1:200 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of PC-12 using 83259-1-RR

  Immunofluorescent analysis of (4% PFA) fixed PC-12 cells using SNAP antibody (83259-1-RR, Clone: 240069A1 ) at dilution of 1:250 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) (SA00013-2).

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  FC experiment of PC-12 using 83259-1-RR

  1x10^6 PC-12 cells were intracellularly stained with 0.25 ug SNAP25 Recombinant antibody (83259-1-RR, Clone:240069A1) and APC-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L)(red), or 0.25 ug Isotype Control (blue). Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

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  Affinity and Kinetic Characterization of 83259-1-RR

  Biolayer interferometry (BLl) kinetic assays of 83259-1-RR against Human SNAP25 were performed. The affinity constant is 4.73 nM.

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Tested Applications

• Positive IHC detected in: mouse brain tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF/ICC detected in: PC-12 cells
• Positive FC (Intra) detected in: PC-12 cells

Recommended dilution

• Immunohistochemistry (IHC): IHC : 1:50-1:500
• Immunofluorescence (IF)/ICC: IF/ICC : 1:125-1:500
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.25 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Product Information

83259-1-RR targets SNAP25 in IHC, IF/ICC, FC (Intra), ELISA applications and shows reactivity with rat samples.

• Tested Reactivity: rat
• Host / Isotype: Rabbit / IgG
• Class: Recombinant
• Type: Antibody
• Immunogen: SNAP25 fusion protein Ag6695
• Full Name: synaptosomal-associated protein, 25kDa
• Calculated Molecular Weight: 23 kDa
• GenBank Accession Number: BC010647
• Gene Symbol: SNAP25
• Gene ID (NCBI): 6616
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

The synaptosomal associated protein of 25 kD (SNAP-25) was first identified as a major synaptic protein by Wilson and colleagues. The protein interacts with syntaxin and synaptobrevin through its N-terminal and C-terminal -helical domains. Its palmitoylation domain is located in the middle of the molecule that contains four cysteine residues. Mutation of the cysteines abolishes palmitoylation and membrane binding. Several elegant studies using synaptosome preparations and permeabilized PC12 cells have suggested that SNAP-25 may act in the late post-docking steps of exocytosis. By limited proteolysis and in vitro binding assay, it is proposed that the two helix domains act independently and contribute equally to form the SNARE complex with syntaxin and synaptobrevin. It seems that a major regulatory element is located in the C-terminus of SNAP-25. Removing a 9 amino acid sequence of SNAP-25 inhibited neurosecretion in chromaffin cells. In addition, it has been shown that inhibition of neurosecretion by AX type E can be rescued by a SNAP-25 C-terminal peptide, probably by initiating the formation of a fusion competent SNARE complex.