N ethylmaleimide sensitive factor attachment protein alpha antibody
N-ethylmaleimide-sensitive factor attachment protein alpha antibody
NSF attachment protein alpha antibody
SNAP alpha antibody
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
IHC image of ab16391 staining in normal human skin formalin fixed paraffin embedded tissue section, performed on a Leica BondTM 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.
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.
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.
References for Anti-Alpha SNAP antibody [4E4] (ab16391)
This product has been referenced in:
Naydenov NG et al. A membrane fusion protein aSNAP is a novel regulator of epithelial apical junctions. PLoS One7:e34320 (2012).
Read more (PubMed: 22485163) »
Wu ZZ et al. Latent membrane protein 1 of Epstein-Barr virus sensitizes cancer cells to cisplatin by enhancing NF-?B p50 homodimer formation and downregulating NAPA expression. Biochem Pharmacol82:1860-72 (2011).
Read more (PubMed: 21945668) »