Alexa Fluor® 594 Anti-JAK2 (phospho Y1007 + Y1008) + JAK1 (phospho Y1034 + Y1035) antibody [E132]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 594 Anti-JAK2 (phospho Y1007 + Y1008) + JAK1 (phospho Y1034 + Y1035) antibody [E132] (AB201744)

ab201744 staining JAK2 (phospho Y1007 + Y1008) in Jurkat cells, starved of serum for 16 hours then treated with 1mM Pervanadate for 30mins at 37°C (Treated) or solvent-only for control purposes (Non-treated). The cells were fixed with 4% formaldehyde (10 min) permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 10% normal goat serum in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab201744 at 1/100 dilution (shown in red) and ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 488) at 1/200 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• WB

Lab

Western blot - Alexa Fluor® 594 Anti-JAK2 (phospho Y1007 + Y1008) + JAK1 (phospho Y1034 + Y1035) antibody [E132] (AB201744)

This data was developed using the same antibody clone in a different buffer formulation (ab32101).

Blocking and dilution buffer : 5% NFDM /TBST.

All lanes:

Western blot - Anti-JAK2 (phospho Y1007 + Y1008) + JAK1 (phospho Y1034 + Y1035) antibody [E132] (<a href='/en-us/products/primary-antibodies/jak2-phospho-y1007-y1008-jak1-phospho-y1034-y1035-antibody-e132-ab32101'>ab32101</a>) at 1/1000 dilution

Lane 1:

293T cells transfected with a Human JAK1 expression vector containing a His tag whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a Human JAK1 expression vector containing a His tag were treated with 50uM pervanadate for 30 min whole cell lysate at 20 µg

Lane 3:

293T cells transfected with a Human JAK1 (mutated Y1034A, mutated Y1035A) expression vector containing a His tag whole cell lysate at 20 µg

Lane 4:

293T cells transfected with a Human JAK1 (mutated Y1034A, mutated Y1035A) expression vector containing a His tag were treated with 50uM pervanadate for 30 min whole cell lysate at 20 µg

Lane 5:

293T cells transfected with a Human JAK2 expression vector containing a His tag whole cell lysate at 20 µg

Lane 6:

293T cells transfected with a Human JAK2 expression vector containing a His tag were treated with 50uM pervanadate for 30 min whole cell lysate at 20 µg

Lane 7:

293T cells transfected with a Human JAK2 (mutated Y1007A, mutated Y1008A) expression vector containing a His tag whole cell lysate at 20 µg

Lane 8:

293T cells transfected with a Human JAK2 (mutated Y1007A, mutated Y1008A) expression vector containing a His tag were treated with 50uM pervanadate for 30 min whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 130 kDa

false

Exposure time: 7s