Immunohistochemistry was performed as previously described.23 (link) Paraffin-embedded tissues were sectioned in a coronal plane at 10–20 μm. The tissue sections were rehydrated, blocked with blocking solution (1% H2O2), and incubated with p-Tau (S202/T205) (1:200), PP2B/PPP3CA (1:200; SantaCruz Biotech), p-Tau (S199) (1:200; Abcam) and anti-III tubulin antibody (1:500 dilutions; Sigma) for 24 h. After washing three times, the slides were processed with Vector ABC Kit (Vector Lab). The immunoreactive signals were developed with DAB chromogen (Thermo Fisher Scientific, Meridian, Rockford, IL, USA) and analyzed under a bright field microscope.
P tau s199
Manufactured by Abcam
Sourced in United States
P-Tau (S199) is a protein marker that detects phosphorylated tau (S199) in biological samples. It is used for research purposes to study tau protein phosphorylation, which is associated with neurodegenerative diseases.
Automatically generated - may contain errors
Lab products found in correlation
Vector abc kit, by Vector Laboratories (3 mentions)
Pp2b ppp3ca, by Santa Cruz Biotechnology (2 mentions)
Dab chromogen, by Thermo Fisher Scientific (1 mentions)
Fitc conjugated secondary antibody, by Vector Laboratories (1 mentions)
Cy3 conjugated secondary antibody, by Jackson ImmunoResearch (1 mentions)
4 protocols using p tau s199
1
Immunohistochemical Analysis of Tau Phosphorylation
2
Immunostaining and Confocal Microscopy of Tau and PP2B Proteins
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Tissues and cells were immunostained for p-Tau (S199) (rabbit polyclonal, 1:200; Abcam, Cambridge, UK), p-Tau (S202/T205) (mouse monoclonal, 1:500; Thermo Scientific, Waltham, MA, USA) and PP2B/PPP3CA (rabbit polyclonal, 1:200; SantaCruz Biotech, Dallas, TX, USA) according to a previous report.22 (link) For confocal microscopy, the specimens were incubated for 1 h with fluorescence (FITC)-conjugated secondary antibody (Vector, Burlingame, CA, USA) and Cy3-conjugated secondary antibody (Jackson Lab, West Grove, PA, USA) after the primary antibody incubation. The images were analyzed using a Spinning Disk Confocal microscope (IX2-DSU, Olympus, Tokyo, Japan). Pre-absorption with excess target protein or omission of primary antibody was used to demonstrate antibody specificity and determine the background generated from the detection assay.
3
Immunohistochemical Analysis of Tau Phosphorylation
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Immunohistochemistry was performed as previously described [22] (link). Animals were deeply anesthetized with 2% avertine (23 μg/g, i.p.) and then perfused through the ascending aorta with 100 ml of cold 0.1 M phosphate buffer saline (PBS), followed by 100 ml of 4% paraformaldehyde in 0.1 M PBS. The brain was removed, post-xed overnight and cryoprotected in 30% sucrose in 0.1 M PBS at 4°C. Then cut by cryotome into 30 μm coronal sections, and processed histochemically as free-oating sections. Tissue sections were then rehydrated, blocked with blocking solution (3% H2O2), and incubated with p-Tau (S202/T205) (1:200; Thermo sher, Waltham, MA, USA) and p-Tau (S199) (1:200; Abcam, Cambridge, MA, USA) for 24 hrs. After three times of washing, the slides were processed with Vector ABC Kit (Vector Lab, Burlingame, CA, USA). Immunoreactive signals were developed with DAB chromogen (Thermo Fisher Scienti c, Waltham, MA, USA). Slides were photomicrographed under bright eld microscopy and analyzed using Image J.
4
Immunohistochemical Analysis of Tau Phosphorylation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Immunohistochemistry was performed as previously described [22] (link). Animals were deeply anesthetized with 2% avertine (23 μg/g, i.p.) and then perfused through the ascending aorta with 100 ml of cold 0.1 M phosphate buffer saline (PBS), followed by 100 ml of 4% paraformaldehyde in 0.1 M PBS. The brain was removed, post-xed overnight and cryoprotected in 30% sucrose in 0.1 M PBS at 4°C. Then cut by cryotome into 30 μm coronal sections, and processed histochemically as free-oating sections. Tissue sections were then rehydrated, blocked with blocking solution (3% H2O2), and incubated with p-Tau (S202/T205) (1:200; Thermo sher, Waltham, MA, USA) and p-Tau (S199) (1:200; Abcam, Cambridge, MA, USA) for 24 hrs. After three times of washing, the slides were processed with Vector ABC Kit (Vector Lab, Burlingame, CA, USA). Immunoreactive signals were developed with DAB chromogen (Thermo Fisher Scienti c, Waltham, MA, USA). Slides were photomicrographed under bright eld microscopy and analyzed using Image J.
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