Anti-JNK1+JNK2 (phospho T183 + Y185)抗体(ab4821)

Proteins were transferred to a nitrocellulose membrane and blocked with 5% skim milk for 1 hour at room temperature.

MEF1 cells were incubated at 37°C for 48h with vehicle control (0 µM) and 5 µM of glibenclamide (ab120267) in DMSO. Increased expression of of JNK1+JNK2 (phospho T183 + Y185) (ab4821) correlates with an increase in glibenclamide concentration, as described in literature.

Whole cell lysates were prepared with RIPA buffer (containing protease inhibitors and sodium orthovanadate), 10µg of each were loaded on the gel and the WB was run under reducing conditions. After transfer the membrane was blocked for an hour using 3% milk before being incubated with ab4821 at 1/1000 dilution and ab85139 at 1 µg /ml overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP (ab97051) at 1/10000 dilution and visualised using ECL development solution.

ab4821 staining JNK1 + JNK2 (phospho T183 + Y185) in A549 cells (green, panel a) by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% paraformaldehyde, permeabilized with 0.25% Triton X-100 and blocked with 5% BSA for 1 hour at room temperature. Samples were incubated with primary antibody (2ug/ml in 1% BSA) for 3 hours at room temperature. An Alexa Fluor® 488-conjugated Goat anti-rabbit IgG polyclonal was used as the secondary antibody (1/400). Nuclei stained with DAPI (blue, panel b), F-actin stained with Alexa Fluor® 594 Phalloidin (red, panel b) and merged images (panel d).

MCF7cells were incubated at 37°C for 4h with vehicle control (0 µM) and different concentrations of cryptotanshinone (ab120666). Increased expression of JNK1+JNK2 (phospho T183 + Y185) in MCF7 cells correlates with an increase in cryptotanshinone concentration, as described in literature.

Whole cell lysates were prepared with RIPA buffer (containing protease inhibitors and sodium orthovanadate), 10µg of each were loaded on the gel and the WB was run under reducing conditions. After transfer the membrane was blocked for an hour using 5% BSA before being incubated with ab4821 at 1/1000 dilution and ab8227 at 1 µg/ml overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP (ab97051) at 1/10000 dilution and visualised using ECL development solution.

ab4821 staining JNK1+JNK2 (phospho T183 + Y185) in human foreskin fibroblasts by ICC/IF. The cells were fixed in cytoskeletal fixative, permeabilized in 0.5% Triton X-100 and blocked in 2% dillution buffer (2%BSA + 0.1% Triton X-100) for 1 hour at 25°C. The primary antibody was diluted, 1/100 and incubated with sample for 12 hours. An Alexa Fluor® 594 conjugated goat polyclonal to rabbit IgG, diluted 1/250 was used as secondary.

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