Anti-APH1a antibody (ab12104)

Western blot

All lanes : Anti-APH1a antibody (ab12104) at 1/500 dilution

Lane 1 : 20ug human brain lysate
Lane 2 : 20ug mouse brain lysate
Lane 3 : 20ug human brain lysate with APH1a peptide (ab12394) at 1 µg/ml
Lane 4 : 20ug mouse brain lysate with APH1a peptide (ab12394) at 1 µg/ml

Secondary
Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP), pre-adsorbed (ab7090) at 1/5000 dilution

Performed under reducing conditions.

Predicted band size : 30 kDa
Observed band size : 30 kDa
Additional bands at : 31 kDa,75 kDa. We are unsure as to the identity of these extra bands.

Exposure time : 30 seconds

Rabbit polyclonal to APH1a (ab12104) used at 1/500 on: Lane 1: Human Brain Lysate Lane 2: Mouse Brain Lysate Lane 3: Human Brain Lysate + 1µg/ml APH1a blocking peptide (ab12394) Lane 4: Mouse Brain Lysate + 1µg/ml APH1a blocking peptide (ab12394) Secondary antibody : Goat polyclonal to Rabbit IgG (HRP) - pre-adsorbed at 1/5000 (ab7090). Expected molecular weight was 30kDa. The band observed between 20-25kDa is likely to be an isoform of Aph1a. We are unsure as to the identity of the minor band seen at 75kDa. Blot exposure time=30 seconds We do not know the identity of the bands at 21 and 75kDa

Immunohistochemistry (PFA perfusion fixed frozen sections) - APH1a antibody (ab12104)

ab12104 staining APH1a in mouse brain tissue section (cortex) by Immunohistochemistry (PFA perfusion fixed frozen sections). Tissue from 4% PFA perfused animals underwent overnight fixation in 4% paraformaldehyde, cryoprotected in 30% sucrose and cut using cryostat.The primary antibody was diluted, 1/1000 (PBS + 0.3% Triton X100) and incubated with sample for 18 hours at 20°C. An abcam antibody ab60314, Chromeo488 conjugated goat polyclonal to rabbit IgG, diluted 1/1000 was used as secondary. ab12104 produces a very pretty staining in neuronal processes in the cortex and the hippocampus.

This image is courtesy of an Abreview submitted by Dr Sophie Pezet