Anti-Alpha SNAP antibody [4E4] (ab16391)

Overview

• Product nameAnti-Alpha SNAP antibody [4E4]See all Alpha SNAP primary antibodies ...
• Description
  Mouse monoclonal [4E4] to Alpha SNAP
• SpecificityDoes not cross react with beta-SNAP.
• Tested applicationsICC/IF, Flow Cyt, WB, IP, ELISA, IHC-P more details
• Species reactivity
  Reacts with: Mouse, Rat, Cow, Human
  
• Immunogen

  Recombinant full length Human alpha SNAP.

• Positive controlRat kidney, brain and MDBK cells.

Properties

• FormLiquid
• Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
• Storage bufferPreservative: None
  Constituents: 50% Glycerol, 0.15M Sodium chloride, 0.02M Sodium phosphate. pH 7.5
• Concentration information loading...
• PurityIgG fraction
• Purification notesThe antibody was purified to >95% homogeneity by standard chromatographic techniques.
• Clonality Monoclonal
• Clone number4E4
• IsotypeIgG1
• Research Areas
  • Signal Transduction
  • Protein Trafficking
  • Vesicle Transport
  • SNAPs & SNAREs

  • Neuroscience
  • Neurotransmission
  • Secretory Vesicles
  • SNAPs & SNAREs

Applications

Target

• FunctionRequired for vesicular transport between the endoplasmic reticulum and the Golgi apparatus.
• Sequence similaritiesBelongs to the SNAP family.
• Cellular localizationMembrane.

Target information above from: UniProt accession P54920 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
• Database links
  • Entrez Gene: 8775 Human
  • Entrez Gene: 108124 Mouse
  • Entrez Gene: 140673 Rat
  • Omim: 603215 Human
  • SwissProt: P81125 Cow
  • SwissProt: P54920 Human
  • SwissProt: Q9DB05 Mouse
  • SwissProt: P54921 Rat

  • Unigene: 126938 Human
  • Unigene: 727492 Human
  • Unigene: 104540 Mouse
  • Unigene: 431371 Mouse
  • Unigene: 31745 Rat

  see all

• Alternative names
  Alpha soluble NSF attachment protein antibodyAlpha-soluble NSF attachment protein antibodyN ethylmaleimide sensitive factor attachment protein alpha antibody

  N-ethylmaleimide-sensitive factor attachment protein alpha antibodyNAPA antibodySNAA_HUMAN antibodySNAP alpha antibodySNAP alpha antibodySNAP-alpha antibodySNAPA antibody

  see all

Anti-Alpha SNAP antibody [4E4] images

• 

  Western blot - alpha SNAP antibody [4E4] (ab16391)
  All lanes : Anti-Alpha SNAP antibody [4E4] (ab16391) at 1/1000 dilution
  
  Lane 1 : NIH3T3 fibroblast cell line whole cell lysate - fraction density 1032
  Lane 2 : NIH3T3 fibroblast cell line whole cell lysate - fraction density 1051
  Lane 3 : NIH3T3 fibroblast cell line whole cell lysate - fraction density 1081
  Lane 4 : NIH3T3 fibroblast cell line whole cell lysate - fraction density 1113
  Lane 5 : NIH3T3 fibroblast cell line whole cell lysate - fraction density 1138
  Lane 6 : NIH3T3 fibroblast cell line whole cell lysate - fraction density 1154
  Lane 7 : NIH3T3 fibroblast cell line whole cell lysate - fraction density 1174
  Lane 8 : NIH3T3 fibroblast cell line whole cell lysate - fraction density 1200
  
  Secondary
  HRP conjugated sheep anti-mouse IgG
  developed using the ECL technique
  
  Performed under reducing conditions.
  
  Observed band size : 36 kDa (why is the actual band size different from the predicted?)
  
  
  Exposure time : 15 minutes

  See Abreview

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)-alpha SNAP antibody [4E4](ab16391)
  IHC image of ab16391 staining in normal human skin formalin fixed paraffin embedded tissue section, performed on a Leica Bond^TM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab16391, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
  
  For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
  
  
• 

  Immunocytochemistry/ Immunofluorescence - Anti-Alpha SNAP antibody [4E4] (ab16391)
  ICC/IF image of ab16391 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab16391, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96879, DyLight® 488 goat anti-mouse IgG (H+L) used at a 1/250 dilution for 1h.Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
  
  
• 

  Flow Cytometry - Anti-Alpha SNAP antibody [4E4] (ab16391)
  Overlay histogram showing HeLa cells stained with ab16391 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab16391, 1µg/1x10⁶ cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
  
  
• 

  Immunocytochemistry/ Immunofluorescence - Anti-Alpha SNAP antibody [4E4] (ab16391)Image from Naydenov NG et al., PLoS One. 2012;7(4):e34320. Epub 2012 Apr 2. Fig 1.; doi:10.1371/journal.pone.0034320; April 2, 2012, PLoS ONE 7(4): e34320.
  Immunofluorescence analysis of Human T84 cell monolayers, staining Alpha SNAP (red) with ab16391.
  
  Epithelial cell monolayers were fixed/permeabilized in 100% methanol for 20 min at -20°C. Cells were blocked in 1% BSA and incubated with primary antibody for 1 hour. An AlexaFluor®568-conjugated anti-mouse IgG was used as the secondary antibody.