pt231 antibody at180  (Thermo Fisher)

Journal: The Journal of Biological Chemistry

Article Title: Sensitive detection and propagation of brain-derived tau assemblies in HEK293-based wild-type tau seeding assays

doi: 10.1016/j.jbc.2025.108245

Figure Lengend Snippet: HA-0N3R AD HEK293 cells propagate AD-brain-derived tau seeds . A , diagram depicting HEK293 cell line expressing HA-0N3R, which has been clonally selected to retain AD-seeded aggregates (HA-0N3R AD ). B , representative immunofluorescent images of HA-0N3R AD cells at passage 4 (P4), probing with phosphorylation-specific antibodies: AT180, AT8, AT100, pS396, and pS422, and with an amyloid-specific dye, Amytracker 680. sb = 10 μm. C , immunoblot of total cell lysate HA-0N3R AD cells over multiple passages. D , immunoblot of the HA-0N3R AD cells over multiple passages using a C-terminal tau antibody (BR134), anti-HA antibody, and phosphorylation-specific antibodies: AT8. AT100, pS396, pS422. Sarkosyl-insoluble extract from approximately 300,000 cells was loaded into each lane.

Article Snippet: The following primary antibodies were used for immunoblotting: Tau 12 (Sigma-Aldrich, MAB2241, 1:2000); BR134 (in-house, rabbit polyclonal, 1:4000); RD3 (Sigma-Aldrich, 05–803, 1:1000); Tau 4R (Cell Signaling Technology; 30,328); anti-GFP (Proteintech, 50430-2-AP, 1:2000); anti-HA.11 (Biolegend, 901516l; 1:2000); anti-FLAG (Sigma-Aldrich, F7425, 1:2000); anti-GAPDH (ThermoScientific, MA5-15738, 1:2000 or Sigma-Aldrich, MAB374, 1:5000); anti-CypA (Bio-techne; AF3589;1:1000); AT100 (pT212/pS214) (Invitrogen, MN1060, 1:500); AT8 (pS202/T205) (Invitrogen, MN1020, 1:500); AT180 (pT231) (Invitrogen, MN0140, 1:500); anti-pS396 (Novus Biologicals, NB100–82243, 1:500); anti-pS422 (Sigma, SAB4300215, 1:500).

Techniques: Derivative Assay, Expressing, Phospho-proteomics, Western Blot

Journal: Frontiers in Neuroanatomy

Article Title: Hibernation Impairs Odor Discrimination – Implications for Alzheimer’s Disease

doi: 10.3389/fnana.2019.00069

Figure Lengend Snippet: List of primary antibodies used on immunoblots of protein fractions.

Article Snippet: AT180 [pT231/pS235] , mouse, mc , 1:500 , Pierce; MN1040 , A.

Techniques: Western Blot

Journal: Frontiers in Neuroanatomy

Article Title: Hibernation Impairs Odor Discrimination – Implications for Alzheimer’s Disease

doi: 10.3389/fnana.2019.00069

Figure Lengend Snippet: Immunohistochemical detection of phospho-tau by the monoclonal antibody AT8in the olfactory bulb of hamsters during hibernation (A) and humans (C) . For the human cases, higher magnification images can be found in the . Legend: gl, glomerular layer; ep, external plexiform layer; m, mitral cell layer; gr, granule cell; lm, stratum lacunosum moleculare; r, stratum radiatum; p, pyramidal cell layer; o, stratum oriens. Scale bars in μm. A schematic description of the distribution of phosphorylated tau is shown for olfactory bulb (B) . Structures containing phosphorylated tau are drawn black whereas those devoid of phosphorylated tau are depicted in light gray. Within the olfactory bulb, olfactory receptor cell axons (oa) are forming synapses to the apical dendritic tuft of mitral cells, which project outside the olfactory bulb. Activity of periglomerular cells (pg), lateral inhibition by granule cells (gc) and efferent modulatory input (e) influence this transmission. Phosphorylation of tau protein in the olfactory bulb was compared in euthermic animals (EU), early torpor (ET), late torpor (TL), and after early arousal (AE) and late arousal (AL). Immunoblots (D) were probed for the presence of specific PHF-like tau-phospho-epitopes (AT 270; AT8; AT 100; AT180; PHF-1), for tau, unphosphorylated at Ser198, Ser199, and Ser202 (Tau-1) and total tau (pan-tau). ANOVA showed significant differences after quantification (E) for AT8 ( p = 0.017), AT100 ( p = 0.00032), AT180 ( p = 2.5e-08), and PHF-1 ( p = 6.5e-08), but not for AT270 ( p = 0.096), Tau-1 ( p = 0.34), and pan-Tau ( p = 0.21). The antibody epitope is indicated below each blot. Inside the diagrams the following significance code for p -values for ANOVA are shown: 0 < “ ∗∗∗ ” < 0.001 < “ ∗∗ ” < 0.01 < “ ∗ ” 0.05 < “n.s.”. The significance codes for p -values for the post hoc Tukey’s HSD test are shown below the diagrams.

Article Snippet: AT180 [pT231/pS235] , mouse, mc , 1:500 , Pierce; MN1040 , A.

Techniques: Immunohistochemical staining, Activity Assay, Inhibition, Transmission Assay, Phospho-proteomics, Western Blot

Journal: Vaccines

Article Title: Doubly Phosphorylated Peptide Vaccines to Protect Transgenic P301S Mice against Alzheimer’s Disease Like Tau Aggregation

doi: 10.3390/vaccines2030601

Figure Lengend Snippet: Normalized phospho-tau contents in brain homogenates of placebo-treated P301S mice ( n = 5–6, ♦) and P301S mice immunized with Tau 199–208 [pS202/pT205] ( n = 3‑5, ), Tau 209–217 [pT212/pS214] ( n = 5–6, ), and Tau 229–237 [pT231/pS235] ( n = 5–6, ) at 32 weeks (upper row) and 48 weeks (lower row) of age obtained from immunoblots. Shown are the relative ratios of phospho-tau (mAbs AT8, AT100, and AT180) to total tau (mAb Tau5) in percent arranged according to the temporal occurrence of the phospho-tau epitopes (pT231/pS235—early, pT212/pS214—intermediate, pS202/pT205—late).

Article Snippet: For immunohistochemistry and immunoblot analysis the following antibodies from Thermo Scientific (Langenselbold, Germany) were used: mouse anti-human PHF-tau IgG 1κ monoclonal antibody, clone AT8 (Tau[pS202/pT205]), mouse anti-human PHF-tau IgG 1κ monoclonal antibody, clone AT100 (Tau[pT212/pS214]) and mouse anti-human PHF-tau IgG 1κ monoclonal antibody, clone AT180 (Tau[pT231]) [ , , , ].

Techniques: Western Blot