Anti-Torsin A antibody (ab34540)

Western blot - Torsin A antibody (ab34540)

All lanes : Anti-Torsin A antibody (ab34540) at 1 µg/ml

Lane 1 : Marker
Lane 2 : Mouse Brain at 10 µg
Lane 3 : Testis (Mouse) Whole Cell Lysate - normal tissue at 10 µg
Lane 4 : Mouse Heart Mouse Tissue Lysate at 10 µg
Lane 5 : Skeletal Muscle (Mouse) Tissue Lysate (ab29711) at 10 µg
Lane 6 : Mouse Fetal lysate - 14 Days old Tissue Lysate at 10 µg
Lane 7 : Brain (Mouse) Tissue Lysate - normal tissue, 0 days old (ab7188) at 10 µg
Lane 8 : Mouse Embryonic Fibroblast 1 at 10 µg
Lane 9 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate (ab7179) at 10 µg

Secondary
IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/15000 dilution

Performed under reducing conditions.

Predicted band size : 38 kDa
Observed band size : 38 kDa
Additional bands at : 105 kDa,65 kDa. We are unsure as to the identity of these extra bands.

Western blot - Torsin A antibody (ab34540)

All lanes : Anti-Torsin A antibody (ab34540) at 1 µg/ml

Lane 1 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate (ab7179)
Lane 2 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
Lane 3 : Liver (Mouse) Tissue Lysate - normal tissue
Lane 4 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate
Lane 5 : Liver (Rat) Tissue Lysate

Lysates/proteins at 10 µg per lane.

Secondary
IRDye 680 Conjugated Goat Anti-Rabbit IgG at 1/10000 dilution

Performed under reducing conditions.

Predicted band size : 38 kDa
Observed band size : 38 kDa
Additional bands at : 65 kDa. We are unsure as to the identity of these extra bands.

Immunocytochemistry/ Immunofluorescence - Torsin A antibody (ab34540)

ICC/IF image of ab34540 stained SHSY5Y cells. The cells were 4% PFA fixed (10 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab34540, 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).

Immunohistochemistry (PFA perfusion fixed frozen sections) - Torsin A antibody (ab34540)

IHC-FoFr image of Torsin A (ab34540) on Rat Cerebral Cortex section. The sections used came from animals perfused fixed with Paraformaldehyde 4%, in phosphate buffer 0.2M. Following postfixation in the same fixative overnight, the brains were cryoprotected in sucrose 30% overnight. Brains were then cut using a cryostat and the immunostainings were performed using the ‘free floating’ technique.

Sophie Pezet, ESPCI, France