Non-treated NIH/3T3, phosphatase inhibitor treated and λ phosphatase treated NIH/3T3 cells were subjected to SDS PAGE followed by western blot with 29223-1-AP (Phospho-S6 Ribosomal protein (Ser235/236) antibody) at dilution of 1:1000 incubated at room temperature for 1 hours.

Non-treated NIH/3T3 and PDGF (HZ-1215) treated NIH/3T3 cells were subjected to SDS PAGE followed by western blot with 29223-1-AP (Phospho-S6 Ribosomal protein (Ser235/236) antibody) at dilution of 1:2000 incubated at room temperature for 1.5 hours.

Non-treated and Calyculin A treated HeLa cells were subjected to SDS PAGE followed by western blot with 67898-1-Ig (Phospho-S6 Ribosomal protein (Ser235) antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with beta tubulin antibody as loading control.

Non-treated and Calyculin A treated HEK-293 cells were subjected to SDS PAGE followed by western blot with 80206-1-RR (Phospho-S6 Ribosomal protein (Ser236) 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.

1X10^6 HEK-293 cells untreated (dashed lines) or treated with Calyculin A (red) were intracellularly stained with 0.5 ug Anti-Human Phospho-S6 Ribosomal protein (Ser236) (80206-1-RR, Clone:7K17) and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) at dilution 1:1000, or 0.5 ug Control Antibody (blue). Cells were fixed with 4% PFA and permeabilized with 90% MeOH.

Serum-free treated NIH/3T3 cells and 10 % fetal bovine serum treated NIH/3T3 cells were subjected to SDS PAGE followed by western blot with 29220-1-AP (Phospho-S6 Ribosomal protein (Ser240/244) antibody) at dilution of 1:3000 incubated at 4℃ overnight. The membrane was stripped and re-blotted with GAPDH antibody as loading control.

Non-treated and ethacrnic acid treated HeLa cells were subjected to SDS PAGE followed by western blot with 80007-1-RR (Phospho-TDP43 (Ser409/410) antibody) at dilution of 1:300 incubated at 4℃ overnight. The membrane was stripped and re-blotted with GAPDH antibody as loading control.

IHC results of Phospho-TDP43 (Ser409/410) rabbit recombinant antibody (80007-1-RR, 1000) with the frontal cortex from FTLD-TDP type B patients. IHC experiment was done with Ventana automatic staining system and Optiview DAB detection kit with heat-induced epitope retrieval (boiling for 32 min in Tris-EDTA based solution CC1 buffer, Ventana). Fig from the lab of Dr. Neumann.

It's human subiculum (high-mag) and temporal cortex (medium-mag) from subject with limbic-predominant age-related TDP-43 encephalopathy neuropathologic change (LATE-NC). Staining provided by Pete Nelson and Ela Patel, U. Kentucky AD Research Center Neuropathology Core.

1X10^6 Jurkat cells untreated (dashed lines) or treated with Calyculin A (red) were intracellularly stained with 0.5 ug Anti-Human Phospho-TDP43 (Ser409/410) (80007-1-RR, Clone:6M10) and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) at dilution 1:1000 , or 0.5 ug Control Antibody (blue). Cells were fixed with 4% PFA and permeabilized with 90% MeOH.

Non-treated HEK-293T and Calyculin A treated HEK-293T cells were subjected to SDS PAGE followed by western blot with 28733-1-AP (ERK1/2-phospho-Thr202/Tyr204) at dilution of 1:3000 incubated at 4℃ overnight. The membrane was stripped and re-blotted with GAPDH antibody as loading control.

Non-treated and Calyculin A treated PC-3 cells were subjected to SDS PAGE followed by western blot with 28733-1-AP (Phospho-ERK1/2 (Thr202/Tyr204) antibody) at dilution of 1:4500 incubated at 4℃ overnight.

IP result of anti-Phospho-ERK1/2 (Thr202/Tyr204)(IP:28733-1-AP, 2ug; Detection:28733-1-AP 1:1000) with Calyculin A treated PC-3 cells lysate 1552 ug.

Non-treated and Calyculin A treated HeLa cells were subjected to SDS PAGE followed by western blot with 67778-1-Ig (Phospho-mTOR (Ser2448) antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

Non-treated and EGF treated NIH/3T3 cells were subjected to SDS PAGE followed by western blot with 67778-1-Ig (Phospho-mTOR (Ser2448) antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

1X10^6 HEK-293 cells were intracellularly stained with 0.2 ug Anti-Human Phospho-mTOR (Ser2448) (67778-1-Ig, Clone:2A12G3) and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L) at dilution 1:1000 (red), or 0.2 ug Control Antibody. Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

Non-treated and Rapamycin treated HEK-293 cells were subjected to SDS PAGE followed by western blot with 67778-1-Ig (Phospho-mTOR (Ser2448) antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with RICTOR antibody (66867-2-Ig) and mTOR antibody (66888-1-Ig) subsequently.

Non-treated and Calyculin A treated HEK-293 cells were subjected to SDS PAGE followed by western blot with 67778-1-Ig (Phospho-mTOR (Ser2448) antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with RICTOR antibody (66867-2-Ig) and mTOR antibody (66888-1-Ig) subsequently.

Immunohistochemical analysis of paraffin-embedded human colon cancer tissue slide using 67778-1-Ig (Phospho-mTOR (Ser2448) antibody) at dilution of 1:1000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

Immunofluorescent analysis of (4% PFA) fixed HepG2 cells using Phospho-mTOR (Ser2448) antibody (67778-1-Ig, Clone: 2A12G3 ) at dilution of 1:200 and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L).

Various lysates were subjected to SDS PAGE followed by western blot with 80024-1-RR (Phospho-JNK (Tyr185) antibody) at dilution of 1:2000 incubated at room temperature for 1.5 hours.