Phospho-Specific Antibodies

ELISA with coating antigen P-tau217 synthetic phosphorylated peptide and 1A9 P-tau217 antibody (TA592983). Negative control: coating Full length tau protein

ELISA with coating antigen P-tau217 synthetic phosphorylated peptide and 1A10 P-tau217 antibody (TA592984). Negative control: coating Full length tau protein

ELISA with coating antigen P-tau217 synthetic phosphorylated peptide and 3B5 P-tau217 antibody (TA592996). Negative control: coating Full length tau protein

ELISA with coating antigen P-tau181 synthetic phosphorylated peptide and 1C12 P-tau181 antibody (TA594030). Negative control: coating Full length tau protein

ELISA with coating antigen P-tau181 synthetic phosphorylated peptide and 5A6 P-tau181 antibody (TA594032). Negative control: coating Full length tau protein

ELISA with coating antigen P-tau181 synthetic phosphorylated peptide and 2F8 P-tau181 antibody (TA594034). Negative control: coating Full length tau protein

Western blot analysis of lysates from HeLa cells

Western blot analysis of lysates from NIH/3T3 cells, using Phospho-STAT3-Y705 Rabbit mAb (TA420055) at 1:1000 dilution. NIH/3T3 cells were treated by TNF-α (20 ng/mL) at 37℃ for 30 minutes.Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) at 1:10000 dilution.Lysates/proteins: 25μg per lane.Blocking buffer: 3% nonfat dry milk in TBST.Detection: ECL Enhanced Kit .Exposure time: 180s.

ELISA with coating antigen P-tau217 synthetic phosphorylated peptide and 1A9 P-tau217 antibody (TA592983). Negative control: coating Full length tau protein

ELISA with coating antigen P-tau217 synthetic phosphorylated peptide and 1A10 P-tau217 antibody (TA592984). Negative control: coating Full length tau protein

ELISA with coating antigen P-tau217 synthetic phosphorylated peptide and 3B5 P-tau217 antibody (TA592996). Negative control: coating Full length tau protein

ELISA with coating antigen P-tau181 synthetic phosphorylated peptide and 1C12 P-tau181 antibody (TA594030). Negative control: coating Full length tau protein

ELISA with coating antigen P-tau181 synthetic phosphorylated peptide and 5A6 P-tau181 antibody (TA594032). Negative control: coating Full length tau protein

ELISA with coating antigen P-tau181 synthetic phosphorylated peptide and 2F8 P-tau181 antibody (TA594034). Negative control: coating Full length tau protein

Western blot analysis of Phospho-AMPK alpha 1/2 (Thr183/Thr172) in various lysates using Phospho-AMPK alpha 1/2 (Thr183/Thr172) antibody.

Western blot using affinity purified anti-Stat2pY690 antibody shows detection of Stat2pY690 protein (arrowhead) in Jurkat cells without (left lane) and with (right lane) 1000U/mL of IFN-a for 15 min at 37oC. Primary antibody was used at 1:1,000.

Western blot analysis of lysates from NIH/3T3 cells

Western blot analysis of lysates from HeLa cells

Western blot analysis of various lysates using Phospho-AKT1-S473 mAb (TA420054) at 1:1000 dilution. Both Jurkat cells and C6 cells were treated by Calyculin A (100 nM) at 37℃ for 30 minutes.Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) at 1:10000 dilution.Lysates/proteins: 25μg per lane.Blocking buffer: 3% nonfat dry milk in TBST.Detection: ECL Basic Kit .Exposure time: 1s.

Western blot analysis of lysates from C6 cells

Phospho specific antibody to Monophosphorylated Regulatory Light Chain of Smooth and Non-muscle Myosin at pS19/pS20 was used at a 1:5000 dilution to detect myosin light chain by WB. Either 13 or 20 µl of a mouse cardiac myocyte lysate was loaded on a 4-20% Criterion gel for SDS-PAGE. Samples were either mock-treated or CLA-treated, as indicated. After washing, a 1:5,000 dilution of HRP conjugated Gt-a-Rabbit IgG preceded color development using Amersham's substrate system.

Western blot analysis of lysates from HT-29 cells, using Phospho-p53-S33 Rabbit pAb (A3185). HT-29 cells were treated by Nocodazole (100ng/mL) for 16 hours.Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) at 1:10000 dilution.Lysates/proteins: 25μg per lane.Blocking buffer: 3% BSA.Detection: ECL Basic Kit .Exposure time: 1s.

Western blot analysis of lysates from HEL cells using Phospho-VASP-S157 Rabbit pAb (TA383199) at 1:1000 dilution. HEL cells were treated by Forskolin (20 uM) at 37℃ for 1 hour.Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) at 1:10000 dilution.Lysates/proteins: 25 μg per lane.Blocking buffer: 3% nonfat dry milk in TBST.Detection: ECL Basic Kit .Exposure time: 0.8s.

Western blot analysis of lysates from 293T cells using Phospho-ERK1-T202/Y204 + ERK2-T185/Y187 Rabbit mAb (TA420033) at 1:1000 dilution (upper) or ERK1/2 Rabbit mAb (A4782) at1:3000 dilution (lower) incubated overnight at 4℃. 293T cells were treated by PMA (100 nM) at 37℃ for 30 minutes after serum-starvation overnight. Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) at 1:10000 dilution.Lysates/proteins: 30 μg per lane. Blocking buffer: 3 % nonfat dry milk in TBST. Detection: ECL Basic Kit . Exposure time: 10s.

Western blot analysis of Phospho-p53 (Ser15) in HeLa treated with HU lysates using Phospho-p53 (Ser15) antibody.The lane on the right is blocked with the Phospho- peptide.

Western blot analysis of lysates from 293 cells, using Phospho-RB-S795 Rabbit pAb (TA380747). 293 cells were treated by 10% FBS at 37℃ for 30 minutes after serum-starvation overnight.Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) at 1:10000 dilution.Lysates/proteins: 25μg per lane.Blocking buffer: 3% BSA.

Western blot analysis of lysates from MCF7 cells

Western blot analysis of various lysates using Phospho-S6 Ribosomal Protein (RPS6)-S240/244 Rabbit pAb (TA381118) at 1:1000 dilution or RPS6 antibody (A6058). HeLa and NIH/3T3 cells were treated by 10% FBS at 37℃ for 30 minutes after serum-starvation overnight.Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) at 1:10000 dilution.Lysates/proteins: 25μg per lane.Blocking buffer: 3% BSA.Detection: ECL Basic Kit .Exposure time: 1s.

Western blot analysis of various lysates

Western blot analysis of Phospho-B23 (Thr199) in HepG2 lysates using Phospho-B23 (Thr199) antibody.The lane on the right is blocked with the synthesized peptide.

Western blot analysis of Phospho-PKA alpha/beta/gamma (Thr197) in mouse brain lysates using Phospho-PKA alpha/beta/gamma (Thr197) antibody.The lane on the right is blocked with the synthesized peptide.

Western blot analysis of Phospho-Rb (Ser780) in various lysates using Phospho-Rb (S780) antibody.

Western blot (WB) analysis of p-Rsk-1 (T359/S363) pAb at 1:2000 dilution Lane1:HEK293T whole cell lysate(40ug) Lane2:HEK293T treated with EGF(100ng/ml,30 minutes) whole cell lysate(40ug)

Western blot analysis of Phospho-Histone H3 (Ser10/Thr11) in 3T3 lysates using Phospho-Histone H3 (Ser10/Thr11)antibody.

Western blot analysis of Phospho-HSF1 (Ser326) in K562, Hela lysates using Phospho-HSF1 (Ser326) antibody.

Immunofluorescent analysis of Jurkat cells treated with Pervanadate, using Phospho-JAK2 (Y1007 + Y1008) Antibody.

Immunofluorescent analysis of HepG2 cells, using Phospho-Nrf2 (S40) Antibody.

Immunofluorescent analysis of MCF7 cells, using Phospho-POLR2A (S2) Antibody .

Western blot analysis of lysates from HeLa cells

Western blot using affinity purified anti-Smad3 pS423 pS425 antibody shows detection of endogenous Smad3 in stimulated cell lysates. Lysates were prepared from control cells (- lanes), or cells stimulated with 2 ng/ml TGF (+lanes) for 1 hour. This reagent recognizes phosphorylated Smad3 and has negligible reactivity against non-phosphorylated Smad3 protein. Personal Communication. Ying Zhang, NIH, CCR, Bethesda, MD.

Western blot analysis of lysates from NIH/3T3 cells

Western blot analysis of various lysates using Phospho-POLR2A CTD-S5 Rabbit pAb (TA380138) at 1:1000 dilution. MCF7 and C2C12 cells were treated by CIP(20uL/400ul) at 37℃ for 1 hour.Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) at 1:10000 dilution.Lysates/proteins: 25μg per lane.Blocking buffer: 3% nonfat dry milk in TBST.Detection: ECL Basic Kit .Exposure time: 1s.

Western blot analysis of lysates from PC-12 cells

Western blot analysis of lysates from NIH/3T3 cells

Western blot analysis of lysates from NIH/3T3 cells

Western blot analysis of lysates from NIH/3T3 cells

Western blot analysis of Phospho-FGFR1 (Tyr654) in Hela lysates using Phospho-FGFR1 (Tyr654) antibody.

Western blot analysis of Phospho-IKK beta (Tyr188) in COS7 lysates using Phospho-IKK beta (Tyr188) antibody.The lane on the right is blocked with the synthesized peptide.

Western blot analysis of Phospho-JAK3 (Tyr785) in 453 lysates using Phospho-JAK3 (Tyr785) antibody.

Western blot analysis of Phospho-kappa Opioid Receptor (Ser369) in various lysates using Phospho-kappa Opioid Receptor (Ser369) antibody.