Mn1050

Manufactured by Thermo Fisher Scientific

Sourced in United States

The MN1050 is a laboratory centrifuge designed for general purpose applications. It can accommodate sample volumes up to 100 mL and can achieve a maximum speed of 6,000 rpm. The MN1050 features a compact and robust construction for reliable performance.

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Lab products found in correlation

T46 antibody against total tau, by Thermo Fisher Scientific (2 mentions) Sector s 600 imager, by MSD (2 mentions) Mn1020, by Thermo Fisher Scientific (2 mentions) Mn1040, by Thermo Fisher Scientific (2 mentions) Tau12, by BioLegend (2 mentions) Paramagnetic beads, by Quanterix (2 mentions) Abn241, by Merck Group (1 mentions) Mab374, by Merck Group (1 mentions) Mn1000, by Thermo Fisher Scientific (1 mentions) H6908, by Merck Group (1 mentions) A 21311, by Thermo Fisher Scientific (1 mentions) Ahb0042, by Thermo Fisher Scientific (1 mentions) F1804, by Merck Group (1 mentions) Blocker a, by MSD (1 mentions) Simoa hd x analyzer, by Quanterix (1 mentions) Simoa hd x, by Quanterix (1 mentions) Neurology 3 plex a assay kit, by Quanterix (1 mentions) Simoa hd 1, by Quanterix (1 mentions)

7 protocols using mn1050

Comprehensive Antibody Characterization Protocol

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Monoclonal antibodies used included: AT270 (MN1050, Thermo Fisher Scientific, RRID: AB_223651), AT180 (MN1040, Thermo Fisher Scientific, RRID: AB_223649), HT7 (MN1000, Thermo Fisher Scientific, RRID: AB_2314654), Tau5 (AHB0042, Thermo Fisher Scientific, RRID: AB_2536235), MAb359 (from Dr. Li Gan), anti-GAPDH (MAB374, Sigma-Aldrich, RRID: AB_2107445), anti-FLAG (F1804, Sigma-Aldrich, RRID: AB_262044), AT8 (MN1020, Thermo Fisher Scientific, RRID: AB223647), anti-SV2 (University of Iowa DSHB, RRID: AB_2315387), anti-PICK1 (75–040, Antibodies Inc, RRID: AB_2164544), anti-KIBRA (sc-133374, Santa Cruz Biotechnology, RRID:AB_2216359). Polyclonal antibodies used included: anti-PKMζ (from Dr. Todd C. Sacktor), anti-PSD95 (2507, Cell Signaling Technology, RRID: AB_561221), anti-HA (H6908, Sigma-Aldrich, AB_260070), GFP (A-21311, Thermo Fisher Scientific, RRID: AB_221477), anti-GluA1 (ABN241, Millipore, RRID: AB_2721164)

Kauwe G., Pareja-Navarro K.A., Yao L., Chen J.H., Wong I., Saloner R., Cifuentes H., Nana A.L., Shah S., Li Y., Le D., Spina S., Grinberg L.T., Seeley W.W., Kramer J.H., Sacktor T.C., Schilling B., Gan L., Casaletto K.B, & Tracy T.E. (2023). KIBRA repairs synaptic plasticity and promotes resilience to tauopathy-related memory loss. bioRxiv.

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Quantifying Vitreous Biomarkers for Neuroinflammation

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Vitreous samples were centrifuged for 15 min at 12,000 rpm to separate the cellular contents, aliquoted at 100 μl, frozen at − 80 °C, and then used for Aβ, t-tau, and p-tau181 measurements. Briefly, assays were run per the manufacturer’s instructions and in duplicate for beta-amyloid 1–40 and 1–42 (Meso Scale Discovery (MSD), Rockville, MD, #K15200E-2), tau phosphorylated at threonine 181 (p-tau181), and t-tau (MSD #K15121D-2), using capturing antibody AT270 against p-tau181 (Thermo Scientific #MN1050) and T46 antibody against total tau as the detecting antibody (Thermo Scientific). Neuroinflammatory cytokines were measured using Neuroinflammation Panel 1 (K15210G, MSD). Samples were diluted 1:2 for pro-inflammatory panel 1 (IFN-γ, IL-10, IL-12p70, IL-13, IL-1β, IL-2, IL-4, IL-6, IL-8, TNF-α), cytokine panel 1 (IL-12/IL-23p40, IL-15, IL-16, IL-17A, IL-1α, IL-5, IL-7, TNF-β, VEGF-A), and angiogenesis panel 1 (basic FGF, VEGFR-1/Flt-1, Tie-2, VEGF-C, VEGF-D), or 1:5 for vascular injury panel 2 (SAA, CRP, VCAM-1, ICAM-1). Sulfo-tag-conjugated anti-mouse secondary antibody (MSD) was used for signal detection by the MSD platform, and an MSD SECTOR S 600 Imager was used to measure the analyte levels.

Subramanian M.L., Vig V., Chung J., Fiorello M.G., Xia W., Zetterberg H., Blennow K., Zetterberg M., Shareef F., Siegel N.H., Ness S., Jun G.R, & Stein T.D. (2020). Neurofilament light chain in the vitreous humor of the eye. Alzheimer's Research & Therapy, 12, 111.

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Quantifying Alzheimer's Biomarkers in Vitreous

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Vitreous samples were centrifuged for 15min at 12,000 rpm to separate the cellular contents, aliquoted at 100 μL, and frozen at −80°C. Protein measurements were performed using the multi-detection SPECTOR 6000 Imager (Meso Scale Discovery (MSD), Rockville, MD) for immunoassay. A 100 μL volume of vitreous fluid aliquot was diluted 1:1 with 1% Blocker A (MSD #R3BA 4) in wash buffer for a total volume of 200 μL.
Samples were subsequently spun down at 17,000 g and 4°C for 15 min, and the supernatant was applied to the immunoassay. Assays were run per the manufacturer’s instructions and in duplicate. Standard immunoassay was used for beta-amyloid 1–40 and 1–42 (MSD #K15200E-2) and tau phosphorylated at threonine 181 (pTau181) together with total tau (MSD #K15121D-2), using capturing antibody AT270 against pTau181 (Thermo Scientific #MN1050) and T46 antibody against total tau as the detecting antibody (Thermo Scientific). Sulfo-tag conjugated anti-mouse secondary antibody (MSD) was used for signal detection by the MSD platform, and an MSD SECTOR S 600 Imager was used to measure analyte levels.

Wright L.M., Stein T.D., Jun G., Chung J., McConnell K., Fiorello M., Siegel N., Ness S., Xia W., Turner K.L, & Subramanian M.L. (2019). Association of Cognitive Function with Amyloid-β and Tau Proteins in the Vitreous Humor. Journal of Alzheimer's disease : JAD, 68(4), 1429-1438.

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Quantitative Protein Biomarkers Measurement

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NfL concentrations were measured using the NF-light™ kit on a Single molecule array (Simoa) HD-X Analyzer (Quanterix, Billerica, MA, USA), following the recommendations by the manufacturer. Plasma p-tau181 levels were measured using an in house Simoa assay as previously described [10 (link)]. Briefly, an AT270 mouse monoclonal antibody (MN1050; Invitrogen, Waltham, MA, USA) was coupled to paramagnetic beads (103,207; Quanterix) and used for capture. As the detector, we used the anti-tau mouse monoclonal antibody Tau12 (806,502; BioLegend, San Diego, CA, USA), conjugated to biotin (A3959; Thermo Fisher Scientific, Waltham, MA, USA), while GSK-3β phosphorylated full-length recombinant tau441 (TO8–50FN; SignalChem, Vancouver, BC, Canada) was used as calibrator. Fluorescent signals were converted to average enzyme per bead numbers as described [19 (link)], and specimen concentrations extrapolated from four-parametric logistic curves generated with known calibrator concentrations.

Clark C., Lewczuk P., Kornhuber J., Richiardi J., Maréchal B., Karikari T.K., Blennow K., Zetterberg H, & Popp J. (2021). Plasma neurofilament light and phosphorylated tau 181 as biomarkers of Alzheimer’s disease pathology and clinical disease progression. Alzheimer's Research & Therapy, 13, 65.

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Immunofluorescence Analysis of Tau Pathology

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Immunofluorescence staining for anti-Tau 5 (1:200, ab3931, Abcam), anti-pTau AT8 (1:200, Cat no. MN1020, Invitrogen), anti-pTau AT180 (1:200, Cat no. MN1040, Invitrogen), anti-pTau PHF-1 (1:200, Cat no. MN1050, Invitrogen), anti-LC3 (1:200, Cat no. M152-3, MBL), anti-P62 (1:200, Cat no. PM045, MBL) and anti-Beclin (1:200, Cat no. PD017, MBL) was performed in Tau-BiFC mice models²⁹. Fluorescence was observed by confocal microscopy (Nikon A1R, JAPAN). Pre-absorption with excess target protein or omission of primary antibody was used to demonstrate antibody specificity and remove background generated by the detection assay. Co-localization and quantitative assessment of images were conducted using NIH Image J v1.44 software.

Subramanian M., Hyeon S.J., Das T., Suh Y.S., Kim Y.K., Lee J.S., Song E.J., Ryu H, & Yu K. (2021). UBE4B, a microRNA-9 target gene, promotes autophagy-mediated Tau degradation. Nature Communications, 12, 3291.

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Plasma p-tau181 Measurement Protocol

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Blood samples were collected and processed according to the ADNI protocol (Kang et al., 2015 (link)). Plasma p-tau181 concentrations were measured at the Clinical Neurochemistry Laboratory, University of Gothenburg (Mölndal, Sweden) using an assay developed in-house on a Simoa HD-X (Quanterix) instrument, as described previously in detail (Karikari et al., 2020 (link)). In brief, the AT270 mouse monoclonal antibody (MN1050; Invitrogen) specific for the threonine-181 phosphorylation site, coupled to paramagnetic beads (103 207; Quanterix) was used for capture and the anti-tau mouse monoclonal antibody Tau12 (806 502; BioLegend), which binds the N-terminal epitope 6-QEFEVMEDHAGT-18 on human tau protein, for detection. All of the available samples were analysed in a single batch. We identified four participants (0.4%) with outlier values of plasma p-tau181 levels that were discarded from subsequent analyses (Supplementary Fig. 1). Longitudinal blood sampling was performed approximately every year, over a median follow-up time of 2.9 years in 938 subjects.

Moscoso A., Grothe M.J., Ashton N.J., Karikari T.K., Rodriguez J.L., Snellman A., Suárez-Calvet M., Zetterberg H., Blennow K, & Schöll M. (2020). Time course of phosphorylated-tau181 in blood across the Alzheimer’s disease spectrum. Brain, 144(1), 325-339.

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Plasma Biomarkers for Neurodegeneration

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Non-fasting blood was drawn from study participants and processed for plasma extraction before storage at -80°C until use. All biomarkers were measured with evaluators blinded to clinical information at the Sahlgrenska Academy at University of Gothenburg in Sweden on the Simoa HD-1 or HD-X platforms (Quanterix, Billerica, MA, USA). Measurements of plasma P-tau181 with the AT270 mouse monoclonal antibody (MN1050; Invitrogen, Waltham, MA, USA) specific for the threonine-181 phosphorylation site was based on an ultrasensitive Simoa immunoassay as described previously-16 with satisfactory cross-site and test-retest reliabilities (Supplementary Data S2). Plasma Aβ42, Aβ40, and total tau (T-tau) were measured using the Neurology 3-plex A assay kit from Quanterix (Billerica, MA, USA). Plasma P-tau181 was not available for six participants, and plasma T-tau, Aβ42, and Aβ40 were not available for three participants due to limited sample availability.

Chong J.R., Ashton N.J., Karikari T.K., Tanaka T., Saridin F.N., Reilhac A., Robins E.G., Nai Y.H., Vrooman H., Hilal S., Zetterberg H., Blennow K., Lai M.K.P, & Chen C.P. (2021). Plasma P-tau181 to Aβ42 ratio is associated with brain amyloid burden and hippocampal atrophy in an Asian cohort of Alzheimer's disease patients with concomitant cerebrovascular disease. Alzheimer's & dementia : the journal of the Alzheimer's Association, 17(10).

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