Anti-PDI antibody - ER Marker (ab31811)

Western blot - PDI antibody (ab31811)

All lanes : Anti-PDI antibody - ER Marker (ab31811) at 1 µg/ml

Lane 1 : Human Brain Whole Tissue Lysate
Lane 2 : Human Kidney Whole Tissue Lysate at 20 µg
Lane 3 : Human Liver Whole Tissue Lysate at 20 µg
Lane 4 : Human Testes Whole Tissue Lysate at 20 µg

Secondary
IR Dye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/15000 dilution

Performed under reducing conditions.

Predicted band size : 57 kDa
Observed band size : 57 kDa


Several smaller bands are seen in Human Liver Whole Tissue Lysate. These are predicted to be break-down products of PDI; the banding pattern is consistent with that observed in Liver lysates for other available PDI antibodies.

- PDI antibody - ER Marker (ab31811)

Image courtesy of Human Protein Atlas

ab31811 staining PDI in Human gall bladder. The paraffin embedded tissue was incubated with ab31811 (1/800 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6. ab31811 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines.

Further results for this antibody can be found at www.proteinatlas.org

Immunocytochemistry/ Immunofluorescence - PDI antibody - ER Marker (ab31811)

ICC/IF image of ab31811 stained human HEK 293 cells. The cells were 4% PFA fixed (10 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab31811, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. The secondary antibody (green) was 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). DAPI was used to stain the cell nuclei (blue). This antibody also gave a positive IF result in HeLa and MCF7 cells.

Western blot - PDI antibody - ER Marker (ab31811)

All lanes : Anti-PDI antibody - ER Marker (ab31811) at 1 µg/ml

Lane 1 : Liver (Mouse) Tissue Lysate
Lane 2 : Liver (Rat) Tissue Lysate

Lysates/proteins at 10 µg per lane.

Secondary
Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP), pre-adsorbed (ab97080) at 1/5000 dilution
developed using the ECL technique

Performed under reducing conditions.

Predicted band size : 57 kDa
Observed band size : 57 kDa


Exposure time : 30 seconds

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PDI antibody - ER Marker (ab31811)

IHC image of ab31811 staining in pancreas formalin fixed paraffin embedded tissue section, performed on a Leica Bond^TM 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 ab31811, 1µg/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.