Anti-UNG antibody (ab23926)

Western blot - UNG antibody (ab23926)

All lanes : Anti-UNG antibody (ab23926) 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 : HeLa (Human epithelial carcinoma cell line) nuclear lysate

Lysates/proteins at 20 µg per lane.

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

Predicted band size : 33, 35 kDa
Observed band size : 35,39 kDa (why is the actual band size different from the predicted?)
Additional bands at : 18 kDa (possible cleavage fragment,cross reactivity),23 kDa (possible cleavage fragment,cross reactivity),40 kDa (possible cross reactivity).

UNG is present in the cell in two isoforms, that differ at their amino-terminal ~40 amino acids. This results in one isoform that is present in the nucleus and one that is present in the mitochondria. This antibody strongly recognizes a doublet at approximately 35 and 39 kDa which closely correspond in size to these two isoforms (whose predicted molecular weights are 33 and 35 kDa).

Immunocytochemistry/ Immunofluorescence - UNG antibody (ab23926)

ICC/IF image of ab23926stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab23926, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iT^TM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - UNG antibody (ab23926)

Image courtesy of Human Protein Atlas

ab23926 staining UNG in human heart muscle. Paraffin embedded human heart muscle tissue was incubated with ab23926 (1/500 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6.

ab23926 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