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Our Abpromise guarantee covers the use of ab2809 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IP||Use at an assay dependent concentration.|
|Inhibition Assay||Use at an assay dependent concentration.|
This antibody detects an ~38 kDa protein representing recombinant rat TGN38.
|Flow Cyt||1/20 - 1/50.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
|ICC/IF||1/20 - 1/50.|
ab2809 staining TGN46 (green) in HepG2 cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formaldehyde, permeabilized with 0.2% Triton X-100 and blocked with 1% donkey serum in 0.1% PBST for 1 hour at 22°C. Samples were incubated with primary antibody (1/50 in blocking solution) for 3 hours at 22°C. An undiluted Alexa Fluor® 488-conjugated donkey anti-mouse IgG polyclonal was used as the secondary antibody. F-actin (blue) was stained using CytoPainter F-actin Staining Kit - Blue Fluorescence (ab112124).
Overlay histogram showing CHO cells stained with ab2809 (red line). The cells were fixed with 100% 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 (ab2809, 1/50 dilution) for 30 min at 22°C. The secondary antibody used was goat anti-mouse DyLight® 488 (IgG H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. 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 CHO cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.
ab2809 staining TGN46 in rat hepatocytescells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formaldehyde, permeabilized with 0.2% Triton X-100 and blocked with 1% donkey serum in 0.1% PBST for 1 hour at 22°C. Samples were incubated with primary antibody (1/50 in blocking solution) for 3 hours at 22°C. An undiluted Alexa Fluor® 488-conjugated donkey anti-mouse IgG polyclonal was used as the secondary antibody.
Blocked with 5% milk in PBST for 1 hour at 22°C.
Incubated with the primary antibody for 12 hours at 4°C in 1% BSA in PBS.
HeLa and COS7 cells were rinsed with PBS and fixed with -20°C methanol for 5 minutes. Cells were incubated with anti-TGN46 antibody (ab2809) diluted 1/100 in PBS containing 0.2% BSA for 1 hour. The cells were then rinsed in PBS and incubated with goat anti-mouse IgG conjugated to rhodamine for 30 minutes in PBS containing 0.2% BSA.
ab2809 staining Chinese Hamster Ovary cell by Immunocytochemistry/Immunofluorescence. The cells were paraformaldehyde fixed and permeabilized in 0.5% Triton prior to blocking with 10% FCS for 30 minutes at room temperature. The primary antibody was diluted 1/1000 and incubated with the sample for 2 hours. The secondary antibody was an Alexa Fluor® 488-conjugated Goat antibody, diluted 1/1000.
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