Anti-Tau (phospho Thr217) Antibody [15B7] (A305063)

Western blot analysis of baculovirus-expressed human Tau 2N4R monomer showing detection of Tau protein phosphorylated at Thr217 using Anti-Tau (phospho Thr217) Antibody [15B7] (A305063) at 1:1,000 for 1 hour at room temperature. Lane 1: Biorad All Blue Standards (3uL), Lane 2: baculovirus expressed hTau 2N4R monomer (2ug), lane 3: baculovirus expressed hTau 2N4R monomer (4ug). Block: 5% Skim Milk in TBST, 1/2 hr at room temperature with shaking. The secondary antibody used was Goat anti-mouse IgG:HRP at 1:4,000 for 1/2 hour at room temperature with shaking. Color Development: Chemiluminescent for HRP (Moss) for 1 min in room temperature. Species reactivity with mouse & Rat. Baculovirus-expressed hTau 2N4R wildtype monomer purified in-house.

Dot blot analysis of Tau 2N4R P301S (phospho & non-phospho) showing detection of Tau protein using Anti-Tau (phospho Thr217) Antibody [15B7] (A305063) at 1:4 for Overnight, 4°C, with shaking. Lane 1: hTau 2N4R P301S Monomer (0.5ug). Lane 2: hTau 2N4R P301S Monomer (1.0ug). Lane 3: Recombinant hTau-441 (2N4R) P301S Monomer, Baculovirus/Sf9 Expressed (phosphorylated) (0.5ug). Lane 4: Recombinant hTau-441 (2N4R) P301S Monomer, Baculovirus/Sf9 Expressed (phosphorylated)(1.0ug). Load: 0.5ug, 1.0ug. Block: 5% BSA in TBST. The secondary antibody used was Goat anti-mouse IgG:HRP at 1:4,000 for 1 hour at room temperature with shaking. Color Development: Chemiluminescent for HRP (Moss) for 5 minutes in room temperature. Dot blot performed with supernatants.

Dot blot analysis of Tau Peptide (pThr217) showing detection of Tau protein using Anti-Tau (phospho Thr217) Antibody [15B7] (A305063) at 1:1,000 for 1 hour at room temperature with shaking. Panel A: hTau 2N4R monomer (pSer214) (2 ug). Panel B: Tau peptide (pThr217) (2 ug). Panel C: Tau peptide (non phospho) (2 ug). Panels on the right are negative control with secondary antibody only. Load: 2ug. Block: 5% Skim Milk in 1X TBST. The secondary antibody used was Goat anti-mouse IgG:HRP at 1:4,000 for 1/2 hour at room temperature with shaking. Color Development: Chemiluminescent for HRP (Moss) for 1 min in ECL. Phospho peptide sequence: SRTPSLP-pT-PPTREPK. Non-phospho peptide sequence: SRTPSLPTPPTREPK. This sequence is the same for human, mouse, and rat.