Non-treated cells and Calyculin A treated cells were subjected to SDS PAGE followed by western blot with 67873-1-Ig (Phospho-MEK1 (Thr292) antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with GAPDH antibody as loading control.
Non-treated HeLa and Calyculin A treated HeLa cells were subjected to SDS PAGE followed by western blot with 67873-1-Ig (Phospho-MEK1 (Thr292) antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with GAPDH antibody as loading control.
Non-treated A431 and Nocodazole treated A431 cells were subjected to SDS PAGE followed by western blot with 67873-1-Ig (Phospho-MEK1 (Thr292) antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with GAPDH antibody as loading control.
Non-treated HEK-293T and Calyculin A treated HEK-293T cells were subjected to SDS PAGE followed by western blot with 80031-1-RR (Phospho-ERK1/2 (Thr202/Tyr204) antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.
Non-treated HeLa and Calyculin A treated HeLa cells were subjected to SDS PAGE followed by western blot with 80031-1-RR (Phospho-ERK1/2 (Thr202/Tyr204) antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.
Non-treated NIH/3T3 and PDGF (HZ-1215) treated NIH/3T3 cells were subjected to SDS PAGE followed by western blot with 80108-1-RR (Phospho-RPS6KA1 (Ser380) antibody) at dilution of 1:2500 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with GAPDH antibody as loading control.
Non-treated HeLa cells and Calyculin A treated HeLa cells were subjected to SDS PAGE followed by western blot with 80108-1-RR (Phospho-RPS6KA1 (Ser380) antibody) at dilution of 1:2500 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with GAPDH antibody as loading control.
UV treated and non-treated NIH/3T3 cells were subjected to SDS PAGE followed by western blot with 80086-1-RR (Phospho-JUN (Ser73) antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with Beta Actin antibody (66009-1-Ig) as loading control.
Non-treated HEK-293T cells, phosphatase inhibitor and λ phosphatase treated HEK-293T cells were subjected to SDS PAGE followed by western blot with 81398-1-RR (Phospho-MNK1 (Thr250/255) antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with GAPDH antibody as loading control.
Immunohistochemical analysis of paraffin-embedded human malignant melanoma tissue slide using 80108-1-RR (Phospho-RPS6KA1 (Ser380) antibody) at dilution of 1:2000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).
Immunohistochemical analysis of paraffin-embedded Jurkat cells slide using 80108-1-RR (Phospho-RPS6KA1 (Ser380) antibody) at dilution of 1:2000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).
1X10^6 HeLa cells untreated (dashed lines) or Calyculin A (red) treated were intracellularly stained with 0.13 ug Anti-Human Phospho-MEK1 (Thr292) (67873-1-Ig, Clone:2D7A8) and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L) at dilution 1:1000, or 0.13 ug Control Antibody (blue). Cells were fixed with 4% PFA and permeabilized with 90% MeOH.
1X10^6 Jurkat cells untreated (dashed line) or treated with TPA treated (red) were intracellularly stained with 0.4 ug Anti-Human Phospho-RPS6KA1 (Ser380) (80108-1-RR, Clone:7F23) and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) at dilution 1:1000 (red), or 0.4 ug Control Antibody (blue). Cells were fixed and permeabilized with True-Nuclear Transcription Factor Buffer Set.
