Anti-U1A antibody (ab40689)

Western blot - U1A antibody (ab40689)

All lanes : Anti-U1A antibody (ab40689) 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
Lane 3 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 4 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
Lane 5 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
Lane 6 : SHSY-5Y (Human neuroblastoma 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 : 31 kDa
Observed band size : 35 kDa (why is the actual band size different from the predicted?)


The U1A protein is acetylated and rich in proline residues (SwissProt data) which may cause it to run at a higher molecular weight than predicted.

Immunocytochemistry/ Immunofluorescence - U1A antibody (ab40689)

ICC/IF image of ab40689 stained human HeLa cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab40689, 5µg/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 - Anti-U1A antibody (ab40689)

Anti-U1A antibody (ab40689) at 1/200 dilution + NSC34 cell nuclear extract at 20 µg

Secondary
IRDye® 800CW Donkey anti-Rabbit IgG at 1/3000 dilution
developed using the ECL technique

Performed under reducing conditions.

Predicted band size : 31 kDa
Observed band size : 31 kDa


Exposure time : 1 minute

This image is courtesy of an Anonymous Abreview.