Anti-Alpha SNAP antibody [4E4]

• Supplier: Abcam
• Catalog: ab16391
• Prices: $380.00
• Sizes: 100 µg
• Host: Mouse
• Clonality: Monoclonal
• Isotype: IgG1
• Clone: 4E4
• Applications: ICC/IF ICC/IF FC WB IP ELISA IHC-P
• Species Reactivities: Mouse, Rat, Bovine, Human
• Antigen: Recombinant full length Human alpha SNAP
• Description: Mouse Monoclonal
• Gene: NAPA
• Conjugate: Unconjugated

Product images

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

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.

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.

Product References

A membrane fusion protein alphaSNAP is a novel regulator of epithelial apical - A membrane fusion protein alphaSNAP is a novel regulator of epithelial apical

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Latent membrane protein 1 of Epstein-Barr virus sensitizes cancer cells to - Latent membrane protein 1 of Epstein-Barr virus sensitizes cancer cells to

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Proteomic changes in rat hippocampus and adrenals following short-term sleep - Proteomic changes in rat hippocampus and adrenals following short-term sleep

Poirrier JE, Guillonneau F, Renaut J, Sergeant K, Luxen A, Maquet P, Leprince P. Proteome Sci. 2008 May 22;6:14.

SNARE proteins mediate fusion between cytosolic lipid droplets and are implicated - SNARE proteins mediate fusion between cytosolic lipid droplets and are implicated

Bostrom P, Andersson L, Rutberg M, Perman J, Lidberg U, Johansson BR, Fernandez-Rodriguez J, Ericson J, Nilsson T, Boren J, Olofsson SO. Nat Cell Biol. 2007 Nov;9(11):1286-93. Epub 2007 Oct 7.