Ps396

Manufactured by Thermo Fisher Scientific

Sourced in United States

The PS396 is a laboratory centrifuge from Thermo Fisher Scientific. It is designed to separate different components of a sample through centrifugal force. The centrifuge can accommodate a range of sample volumes and rotor types, allowing it to be used in various laboratory applications.

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Lab products found in correlation

Total tau, by Thermo Fisher Scientific (1 mentions) Ecl kit, by Cytiva (1 mentions) Protease inhibitor cocktail, by Merck Group (1 mentions) Dc protein assay, by Bio-Rad (1 mentions) Phosphatase inhibitor cocktail, by Roche (1 mentions) Las 1000, by Fujifilm (1 mentions) Nitrocellulose membrane, by Bio-Rad (1 mentions) Anti pink1, by Abcam (1 mentions) Anti mfn2, by Abcam (1 mentions) Mab anti opa1, by BD (1 mentions) Anti cytc, by Abcam (1 mentions) Anti parkin, by Abcam (1 mentions) Anti α tubulin mab, by Merck Group (1 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions) Anti gapdh, by Abcam (1 mentions) Anti gfp, by Abcam (1 mentions) Anti gm130, by BD (1 mentions) Synaptotagmin, by BD (1 mentions) Psd 95, by BD (1 mentions) β tubulin, by Merck Group (1 mentions)

5 protocols using ps396

Quantifying Peripheral Tau Levels

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The levels of total tau in peripheral organs tissues were measured by
enzyme-linked immunosorbent assay (ELISA) Kits (Thermo Fisher Scientific),
following the manufacturer’s instructions. For the second series of
submandibular glands, total tau (Thermo Fisher Scientific), and levels of
phosphorylated tau detected by pT231 (Thermo Fisher Scientific) and pS396
(Thermo Fisher Scientific) were quantified. All steps were performed at room
temperature. Optical density of each well was measured immediately after adding
Stop Solution using a plate reader set at an absorbance of 450 nm for 1 second;
sample concentrations were calculated against the generated standard curves.
total tau and S396 data were then adjusted to ng tau/mg sample protein, while
T231 data were adjusted to T231 units/mg sample protein.

Dugger B.N., Whiteside C.M., Maarouf C.L., Walker D.G., Beach T.G., Sue L.I., Garcia A., Dunckley T., Meechoovet B., Reiman E.M, & Roher A.E. (2016). The presence of select tau species in human peripheral tissues and their relation to Alzheimer’s disease. Journal of Alzheimer's disease : JAD, 51(2), 345-356.

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Tau Protein Quantification in Transgenic Mice

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Three-month-old female P301L human tau transgenic mice (JNPL3), having an average weight of 32 g at the time of testing were used. They received a single dose of SAR502250 (1, 3, 10, 30 and 100 mg/kg/d) by oral route. One hour after the administration, brains and spinal cords were rapidly dissected and quickly frozen. Tissue was homogenized with homogenization buffer (62.5 mM Tris-HCl pH 6.8, 2.3% SDS, 1 mM EDTA, 1 mM EGTA, 1 mM DTT, Protease inhibitor cocktail (Sigma-Aldrich), Phosphatase inhibitor cocktail (Roche Diagnostics). Homogenized sample was boiled for 5 min and centrifuged at 15,000 x g for 15 min. Supernatant was collected and protein concentration was measured by DC protein assay (Bio Rad). 10 μg of samples were applied on 10% SDS-PAGE and transferred onto nitrocellulose membranes. Total human tau protein and phosphorylated (S₃₉₆) tau protein was evaluated by western-blotting labelling with TauN (BD Transduction) and PS396 (Thermo Fisher Scientific) antibodies respectively. Each band was visualized with ECL kit (Amersham Bioscience) and detected with LAS 1000 (Fuji Film).

Griebel G., Stemmelin J., Lopez-Grancha M., Boulay D., Boquet G., Slowinski F., Pichat P., Beeské S., Tanaka S., Mori A., Fujimura M, & Eguchi J. (2019). The selective GSK3 inhibitor, SAR502250, displays neuroprotective activity and attenuates behavioral impairments in models of neuropsychiatric symptoms of Alzheimer’s disease in rodents. Scientific Reports, 9, 18045.

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Quantifying Tau Phosphorylation Patterns

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Cell lysates were resolved via SDS-PAGE (on 10% gels for tau-T40 and 15% gels for tau-K18), transferred to nitrocellulose membrane (Biorad), and blocked with 2% milk in 1X TBS for 30 min. Membranes were incubated with primary antibody overnight at 4 °C, followed by a 1 h room temperature incubation with HRP-conjugated anti-mouse or anti-rabbit antibodies (1:1000). The following primary antibodies were used: pSer202/pThr205-tau (AT8, 1:1000, Thermo Fisher Scientific, MN1020), pSer262-tau (p-S262, 1:1000, Thermo Fisher Scientific, 44-750G), pThr231-tau (AT180, 1:1000, Thermo Fisher Scientific, MN1040), pThr181-tau (AT270, 1:1000, Thermo Fisher Scientific, MN1050), tau-1 (1:1000, Millipore, MAB3420), pSer396-tau (p-S396, Thermo Fisher Scientific, 44-752G), anti-acetylated K280-tau4 (link) (ac-K280, 1:1000), tau K9JA (total tau, 1:5000, Dako, A0024), tau-T46 (1:1000, Thermo Fisher Scientific, 13-6400), and Hsc70 (1:1000, Enzo Life Sciences, SPA-815). ImageJ software (NIH) was used to quantify band intensity.

Trzeciakiewicz H., Tseng J.H., Wander C.M., Madden V., Tripathy A., Yuan C.X, & Cohen T.J. (2017). A Dual Pathogenic Mechanism Links Tau Acetylation to Sporadic Tauopathy. Scientific Reports, 7, 44102.

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Plasmid Construction and Antibody Validation

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Human pIRES-eGFP-Tau40 plasmid was a gift of Dr. Khalid Iqbal (New York State Institute for Basic Research in Developmental Disabilities, Staten Island, NY). The PCI-neo-Tau40 was constructed in our lab. For GFP-parkin (PK2, the full-length mouse parkin cDNA) was cloned into pEFGP-C1 plasmid (Clontech). EGFP-labeled AAV2/8-htau and its control virus were purchased from OBio Biologic Technology Co., Ltd.
Mouse monoclonal antibody (mAb) anti-OPA1 and anti-GM130 were from BD Bioscience; mAb anti-Mfn1 was from Santa Cruz; mAb anti-COXIV, anti-GAPDH, anti-Cytc, anti-GFP, anti-PDI1 and pAb anti-ubiquitin, anti-PINK1, anti-Parkin, anti-Mfn2, anti-SQSTM/p62, anti-LC and anti-TOMM40 were from Abcam; mAb anti-α-tubulin was from Sigma; two kinds of mAb anti-tau-5 were from NeoMarkers or Millopore, mAb tau-1 was from Chemicon, pAbs against phosphorylated tau (pT205, pS214, pS262, pS396 or pS404) were from BioSource; mAb AT8 (tau phosphorylated at Ser202 and Thr205) was from Thermo Fisher Scientific, pAb anti-TOMM20 was from Anbo Biotechnology, CCCP/JC-1 was from Sigma; Lipofectamine2000 and TMRM was from Invitrogen.

Hu Y., Li X.C., Wang Z.H., Luo Y., Zhang X., Liu X.P., Feng Q., Wang Q., Yue Z., Chen Z., Ye K., Wang J.Z, & Liu G.P. (2016). Tau accumulation impairs mitophagy via increasing mitochondrial membrane potential and reducing mitochondrial Parkin. Oncotarget, 7(14), 17356-17368.

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Tau Protein Antibodies for Western Blotting

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E1 (31 (link)), a polyclonal antibody specific to human tau (aa 19-33, unphosphorylated), was prepared in our laboratory. MS06, a polyclonal antibody specific to mouse tau, was raised against mouse tau polypeptide corresponding to amino acid residue 118–131 (SKDRTGNDEKKAKG). Tau5, pS199, pT231, and pS396 were purchased from Biosource International (Camarillo, CA, USA). Tau1 was from Chemicon (Temecula, CA, USA). Monoclonal antibodies to β-actin and β-tubulin were purchased from Sigma. Monoclonal antibodies to GAP-43, PSD-95, and synaptotagmin were purchased from BD Transduction Laboratories (San Jose, CA, USA). For western blotting, antibodies were used at the following dilutions in blocking solution: E1, 1:5,000; MS06, 1:2,000; Tau1, 1:5,000; Tau5, 1:2,000; pS199, 1:5,000; pT231, 1:2,000; pS396, 1:2,000; β-actin, 1:5,000; β-tubulin, 1:5,000; GAP-43, 1:2,000; PSD-95, 1:2,000; synaptotagmin, 1:2,000.

Sahara N., Murayama M., Higuchi M., Suhara T, & Takashima A. (2014). Biochemical Distribution of Tau Protein in Synaptosomal Fraction of Transgenic Mice Expressing Human P301L Tau. Frontiers in Neurology, 5, 26.

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