Western blot - PDI antibody - ER Marker (ab3672)

Anti-PDI antibody - ER Marker (ab3672) at 1/1000 dilution + PC12 whole cell lysate at 20 µg

Secondary
HRP conjugated swine anti-rabbit antibody
developed using the ECL technique

Performed under reducing conditions.

Observed band size : 60 kDa (why is the actual band size different from the predicted?)
Additional bands at : 36 kDa,48 kDa,58 kDa,75 kDa. We are unsure as to the identity of these extra bands.

Exposure time : 2 minutes

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - PDI antibody - ER Marker (ab3672)

ab3672 at 1/400 staining mouse kidney tissue sections by IHC-P. The tissue was formladehyde fixed and a heat mediated antigen retrieval step was performed in citrate buffer. The tissue was incubated with the primary antibody for 30 minutes at 22°C and then an HRP conjugated goat anti-rabbit antibody was used as the secondary.

This image is courtesy of an anonymous Abreview

Immunohistochemistry (Frozen sections) - PDI antibody - ER Marker (ab3672)

ab3672 staining mouse cortical oligodendrocytes from vibratome sections by IHC-Fr.  Sections were fixed in Somogyi (PFA + glutaraldehyde + picric acid) or PLP (Periodate-lysine-PFA) and permeabilized in TX-100, prior to blocking with 2.5% serum for 2 hours at 25°C.  The primary antibody was diluted 1/1000 and incubated with the sample for 18 hours at 25°C.  A TRITC conjugated pig anti-rabbit IgG antibody, diluted 1/1000, was used as the secondary.

This image is courtesy of an Abreview submitted by Dr Guillermo Estivill-Torrus

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - PDI antibody - ER Marker (ab3672)

ab3672 staining PDI - ER in human HaCaT Keratinocyte by ICC/IF. The cells were paraformaldehyde fixed, permeabilized in 0.25% triton X-100 buffer and blocked in 2.5% BSA plus 1% goat serum for 1 hour at 4°C. The primary antibody was diluted 1/100 and incubated with sample for 1 hour at 24°C. An Alexa Fluor® 488 conjugated goat polyclonal to rabbit IgG, diluted 1/1000 was used as secondary.

This image is courtesy of an anonymous Abreview

Immunocytochemistry/ Immunofluorescence - PDI antibody - ER Marker (ab3672)

ICC/IF image of ab3672 stained HeLa cells. The cells were 4% PFA 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 (ab3672, 1æg/ml) overnight at +4øC. The secondary antibody (green)ÿwas Alexa Fluor© 488 goat anti-rabbit IgG (H+L) used at a 1/1000 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 1.43æM.