ICC/IF image of ab13506 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 ab13506 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- mouse (ab96879) 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.
Western blot - ERp57 antibody [MaP.Erp57] (ab13506)This image is courtesy of an Abreview submitted by Pawel Mazur
Predicted band size : 61 kDa Observed band size : 61 kDa
This image is courtesy of an Abreview submitted by Pawel Mazur
ERp57 antibody (ab13506) at 1/1000 dilution (in PBST) for 8 hours at 4°C + lysate of mouse of pancreatic cancer cell line at 25 µg.
An HRP conjugated Sheep anti-mouse IgG polyclonal at 1/5000 dilution developed using the ECL technique
Performed under non-reducing conditions.
Exposure time: 1 minute
Blocking Step: 10% Milk for 1 hour at room temperature.
Immunohistochemistry (Frozen sections) - ERp57 antibody [MaP.Erp57] (ab13506)This image is courtesy of an anonymous Abreview
ab13506 staining ERp57 in Mouse duodenum, pancreas, and pancreatic neoplastic tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with acetone:methanol (1:1), permeabilized with PBS + Triton X-100 (0.025%) and blocked with 10% serum for 1 hour at room temperature. Samples were incubated with primary antibody (1/250 in PBS) for 8 hours at 4°C. An Alexa Fluor®488-conjugated Goat anti-mouse IgG polyclonal (1/500) was used as the secondary antibody. Nuclei were stained by DAPI.
Overlay histogram showing HeLa cells stained with ab13506 (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. The cells were then incubated with the antibody (ab13506, 2µg/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 in 0.1% PBS-Tween used under the same conditions.
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