Anti-PICK1 antibody (ab3420)

Western blot - PICK1 antibody (ab3420)

Lane 1 : An myc antibody in PBST for 16 hours at 4°C
Lane 2 : Anti-PICK1 antibody (ab3420) at 1/500 dilution (in PBST for 16 hours at 4°C)

Lane 1 : Transfected CHO cells expressing human Pick1 tagged with myc
Lane 2 : Transfected CHO cells expressing human Pick1 tagged with myc

Lysates/proteins at 20 µg per lane.

Secondary
Lane 2 : An HRP-conjugated goat anti-rabbit IgG polyclonal at 1/10000 dilution

developed using the ECL technique

Performed under reducing conditions.

Observed band size : ~52 kDa (why is the actual band size different from the predicted?)
Additional bands at : ~40 kDa. We are unsure as to the identity of these extra bands.

Exposure time : 2 minutes

Blocking Step: 5% Milk for 1 hour at 20°C

This image is courtesy of an anonymous Abreview

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - PICK1 antibody (ab3420)

IHC image of ab3420 staining in rat adult brain formalin fixed paraffin embedded tissue section, performed on a Leica Bond^TM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab3420, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

Immunocytochemistry/ Immunofluorescence - PICK1 antibody (ab3420)

ab3420 staining PICK1 in CHO cells stably transfected with Human PICK1 by Immunocytochemistry/ Immunofluorescence. Cells were fixed in paraformaldehyde and permeabilized in 0.5% Triton X-100 prior to blocking in 10% serum for 1 hour at room temperature. The primary antibody was diluted 1/1000 in PBS, 0.5% Triton, 10% FCS and incubated with the sample for 2 hours at ºC. The secondary antibody was Alexa Fluor® 488-conjugated Goat anti-Rabbit polyclonal, diluted 1/500.

This image is courtesy of an Anonymous Abreview.

Western blot - PICK1 antibody (ab3420)

Western blot of PICK1 on AtT20 cell lysate using ab3420.