Anti-JAK2 抗体 [EPR108(2)] - BSA and Azide free (ab170718)

This data was developed using the same antibody clone in a different buffer formulation (ab108596).

  Lanes 1- 4: Merged signal (red and green). Green - ab108596 observed at 131 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

 ab108596 was shown to react with JAK2 in wild-type A549 cells in western blot. Loss of signal was observed when knockout cell line ab267113 (knockout cell lysate ab256963) was used. Wild-type A549 and JAK2 knockout A549 cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab108596 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

ab108596 at 1/500 dilution (3.0µg/ml) immunoprecipitating JAK2 in K562 (Human chronic myelogenous leukemia lymphoblast)  cell lysate.

Lane 1 (input): K562(Human chronic myelogenous leukemia lymphoblast) whole cell lysate 10µg

Lane 2 (+): K562 whole cell lysate, 350µg  + ab108596, 2µg

Lane 3 (-): K562 cell lysate, 350µg  + rabbit IgG (ab172730), 2µg

For western blotting, ab131366 VeriBlot for IP (HRP) was used at 1/1000.

Blocking and dilution buffer: 5% NFDM/TBST.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108596).

Immunocytochemistry/ Immunofluorescence analysis of K562 (Human chronic myelogenous leukemia cell line from bone marrow) cells labeling JAK2 with purified ab108596 at 1/150 dilution (8.5μg/ml). Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, a goat anti-rabbit IgG(Alexa Fluor^® 488) was used as the secondary antibody at 1/1000 dilution. Ab195889, anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) was used as the counter stain at 1/200 (2.5 μg/ml). PBS instead of the primary antibody was the negative control. DAPI was used as a nuclear counterstain.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108596).

Immunocytochemistry/Immunofluorescence analysis of Ramos (Human Burkitt's lymphoma) cells labelling JAK2 with unpurified 108596 at 1/300 (7.0 μg/mL). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000, 2 μg/mL) was used as the secondary antibody. Nuclei were stained with DAPI (blue).

Confocal image showing nuclear and cytoplasmic staining on Ramos cell line.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108596).

Immunocytochemistry/Immunofluorescence analysis of Jurkat (Human T cell leukemia cells from peripheral blood) cells labelling JAK2 with unpurified ab108596 at 1/300 (7.0 μg/mL). Cells were fixed with 4% Paraformaldehydeand permeabilized with 0.1% Triton X-100. ab150077, Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000, 2 μg/mL) was used as the secondary antibody. Nuclei were stained with DAPI (blue).

Confocal image showing nuclear and cytoplasmic staining on Jurkat cell line. 

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108596).