Anti-DNA polymerase alpha antibody (ab31777)

Western blot - DNA polymerase alpha antibody (ab31777)

All lanes : Anti-DNA polymerase alpha antibody (ab31777) at 1 µg/ml

Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate (ab7899)
Lane 3 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate (ab7909)

Lysates/proteins at 20 µg per lane.

Secondary
Goat polyclonal to Rabbit IgG (Alexa Fluor® 680) at 1/10000 dilution

Performed under reducing conditions.

Predicted band size : 166 kDa
Observed band size : 180 kDa (why is the actual band size different from the predicted?)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - DNA polymerase alpha antibody (ab31777)

Image courtesy of Human Protein Atlas ab31777 staining DNA polymerase alpha in human thyroid gland, showing a strong staining in nuclei of the glandular cells. Paraffin embedded human thyroid tissue was incubated with ab31777 (1/100 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6.

ab31777 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 - DNA polymerase alpha antibody (ab31777)

ICC/IF image of ab31777 stained human MCF7 cells. The cells were 4% PFA fixed (10 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab31777, 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, HEK 293 and HepG2 cells.