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 applicationsSuitable for: ICC/IF, Flow Cyt, WB, IP, ELISA, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Cow, Human
  • Immunogen

    Recombinant full length Human alpha SNAP.

  • Positive control
    • Rat kidney, brain and MDBK cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab16391 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF Use a concentration of 5 µg/ml.
Flow Cyt Use 1µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
WB 1/5000. Detects a band of approximately 36 kDa.
IP Use a concentration of 2 - 10 µg/ml.
ELISA Use at an assay dependent concentration.
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Target

Anti-Alpha SNAP antibody [4E4] images

  • 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

  • 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.

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 One 7:e34320 (2012). WB, ICC/IF . 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 Pharmacol 82:1860-72 (2011). WB ; Mouse . Read more (PubMed: 21945668) »

See all 4 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Cell lysate - whole cell (NIH3T3 fibroblast cell line)
Specification NIH3T3 fibroblast cell line
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
Username

Dr. Mikael Rutberg

Verified customer

Submitted Feb 03 2006

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"