Anti-ADP Ribosylation Factor antibody [1D9] (ab2806)

Immunoprecipitation - ADP Ribosylation Factor antibody [1D9] (ab2806)

Ab2806 immunoprecipitating ADP Ribosylation Factor from HeLa cells. 10ul of antibody were added to 500ul of HeLa cell lysate (at 1.5mg/ml) and incubated overnight before precipitation with Protein G. Negative control lysate received the same treatment except no antibody was added. ab2806 was used at 1/500 in the western blotting stage and the secondary antibody was an HRP conjugated anti-mouse IgG. The membrane was finally stained with amido black.

Lane 1: negative control

Lane 2: ab2806

This image is courtesy of an Abreview submitted by Denis Krndija submitted on 5 January 2006.

Western blot - ADP Ribosylation Factor antibody [1D9] (ab2806)

All lanes : Anti-ADP Ribosylation Factor antibody [1D9] (ab2806) at 1/500 dilution

Lane 1 : Lysates prepared from human Huh-7 cells
Lane 2 : Lysates prepared from human Huh-7 cells

Lysates/proteins at 15000 cells per lane.

Secondary
HRP-conjugated sheep polyclonal to mouse IgG at 1/10000 dilution

Performed under reducing conditions.

Observed band size : 21 kDa (why is the actual band size different from the predicted?)


Exposure time : 4 minutes

This image is a courtesy of Anonymous Abreview

Flow Cytometry-ADP Ribosylation Factor antibody [1D9](ab2806)

Overlay histogram showing HeLA cells stained with ab2806 (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 (ab2806, 1/20 dilution) 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.