Anti-Alkyl-DHAP synthase antibody (ab79122)

  Western blot - Alkyl-DHAP synthase antibody (ab79122)

All lanes : Anti-Alkyl-DHAP synthase antibody (ab79122) at 1 µg

Lane 1 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
Lane 2 : U937 (Human leukemic monocyte lymphoma cell line) Whole Cell Lysate
Lane 3 : THP1 (Human acute monocytic leukemia cell line) Whole Cell Lysate
Lane 4 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 5 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate

Lysates/proteins at 10 µg per lane.

Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
developed using the ECL technique

Performed under reducing conditions.

Predicted band size : 73 kDa
Observed band size : 72 kDa (why is the actual band size different from the predicted?)
Additional bands at : 18 kDa. We are unsure as to the identity of these extra bands.

Exposure time : 1 minute

  Immunocytochemistry/ Immunofluorescence - Alkyl-DHAP synthase antibody (ab79122)

ICC/IF image of ab79122 stained HeLa cells. The cells were 100% Methanol fixed (5 min) and then incubated in 1%BSA / 10% normal Goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab79122, 1µg/ml) overnight at +4°C. 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) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 100% Methanol fixed (5 min) HEK293 cells at 1µg/ml, and in 4% PFA fixed (10 min) HeLa and HEK293 cells at 1µg/ml.