Anti-ABCE1 antibody (ab32270)

Western blot - ABCE1 antibody (ab32270)

All lanes : Anti-ABCE1 antibody (ab32270) at 1 µg/ml

Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 20
Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate (ab7899) at 20 µg
Lane 3 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate (ab7909) at 20 µg

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

Performed under reducing conditions.

Predicted band size : 67 kDa
Observed band size : 70 kDa (why is the actual band size different from the predicted?)
Additional bands at : 100 kDa. We are unsure as to the identity of these extra bands.

Immunocytochemistry/ Immunofluorescence - ABCE1 antibody (ab32270)

ICC/IF image of ab32270 stained human HeLa cells. The cells were methanol fixed (5 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab32270, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal serum / 0.3M glycine was used to quench autofluorescence and 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).

Western blot - ABCE1 antibody (ab32270)

All lanes : Anti-ABCE1 antibody (ab32270) at 1/250 dilution

Lane 1 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
Lane 2 : Testis (Mouse) Tissue Lysate - normal tissue
Lane 3 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate

Lysates/proteins at 10 µg per lane.

Secondary
IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

Performed under reducing conditions.

Predicted band size : 67 kDa
Observed band size : 67,70 kDa (why is the actual band size different from the predicted?)
Additional bands at : 110 kDa. We are unsure as to the identity of these extra bands.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - ABCE1 antibody (ab32270)

Image courtesy of Human Protein Atlas ab32270 staining ABCE1 in human duodenum, showing a distinct cytoplasmic staining pattern. Paraffin embedded human duodenum tissue was incubated with ab32270 (1/300 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6.

ab32270 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues. Further results for this antibody can be found at www.proteinatlas.org