Pa1 183

Manufactured by Thermo Fisher Scientific

The PA1-183 is a laboratory equipment designed for general laboratory use. It is a precision instrument that can perform various tasks required in a research or analytical setting. The core function of the PA1-183 is to provide accurate and reliable measurements, but a detailed description of its intended use is not available.

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Lab products found in correlation

6 protocols using pa1 183

Western Blot Analysis of Caudal Disc Proteins

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Protein from caudal discs of seven mice were extracted using Tissue Protein Extraction Reagent (T‐PER) with proteinase inhibitor cocktail per the manufacturer's instructions (Thermo Fisher 78510, Waltham, MA). Western blotting for the expression of p53, matrix metalloproteinase‐3 (MMP‐3), and a disintegrin and metalloproteinase with thrombospondin motifs‐4 (ADAMTS‐4) was also performed. Primary antibodies (1:1000 dilution) to p53 (Cell Signaling Technology 2524, Danvers, MA), MMP‐3 (Abcam ab52915, Cambridge, United Kingdom), ADAMTS‐4 (Abcam ab185722, Cambridge, United Kingdom), and β‐actin (Thermo Fisher PA1‐183, Waltham, MA) and secondary anti‐rabbit horse radish peroxidase antibody (1:5000 dilution, Thermo Fisher 31460, Waltham, MA) were used.
Following electrophoresis on Tris‐HEPES 4%‐20% gradient polyacrylamide denaturing gel (Thermo Scientific 25204, Waltham, MA), separated proteins were transferred to a polyvinylidene difluoride membrane. Signals were measured using chemiluminescent detection (Thermo Scientific 34096, Waltham, MA) and ChemiDoc MP system (Bio‐Rad, Hercules, CA). The protein bands were analyzed using NIH Image J 1.44p with local background subtraction. The data obtained were normalized with β‐actin.

Kritschil R., Zhang Z., Lei C., Zhong J., Dong Q., Lee J., Conover C.A., Sowa G., Vallejo A.N, & Vo N. (2020). Effects of suppressing bioavailability of insulin‐like growth factor on age‐associated intervertebral disc degeneration. JOR Spine, 3(4), e1112.

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Comprehensive α-Synuclein and Neurodegeneration Analysis

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Human specific α-synuclein, clone Syn 211 (αSyn) (1:500, Thermo Fisher #AHB0261); anti-phosphorylated α-synuclein at serine 129 [pSyn (S129)] (1:1000, Abcam #ab51253); phosphorylated α-synuclein at tyrosine 125 (pSyn Y125) (1:250, Abcam #ab10789); anti-glial fibrillary acidic protein (GFAP) (1:2000, Abcam #ab4674); anti-glutamate aspartate transporter (GLAST) (1:250, Miltenyi Biotec #130-095-822); anti-ubiquitin (1:500, Thermo Fisher #13-1600); anti-p62/sequestosome 1 (p62) (1:1000, Abcam #ab56416); anti-lysosomal-associated membrane protein 1 (LAMP1) (1:1000, Abcam #ab25245); anti-microtubule associated protein 2 (Map 2) (1:5000, Abcam #ab5392); III β-tubulin (1:1000, Neuromics #MO15013); anti-ionized calcium binding adaptor molecule 1 (Iba1) (1:500, Wako, #019-19,741); β-actin loading control (1:5000, Thermo Fisher #PA1-183); the (E,E)-1-Fluoro-2,5-bis(3-hydroxycarbonyl-4-hydroxy) styrylbenzene (FSB dye) (Congo red derivative) (at 2.5 μM, Santa Cruz #760988-03-2).

Krejciova Z., Carlson G.A., Giles K, & Prusiner S.B. (2019). Replication of multiple system atrophy prions in primary astrocyte cultures from transgenic mice expressing human α-synuclein. Acta Neuropathologica Communications, 7, 81.

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Western Blot Analysis of Raphe Nuclei Proteins

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The protein expression levels of Iba-1, C1q, cleaved caspase 3, synaptophysin, PSD95, and β-actin in raphe nuclei homogenates were evaluated using Western blotting. The protein concentrations of the homogenates were equalized, and the samples were separated by 10% polyacrylamide gel electrophoresis and transferred to polyvinylidene fluoride membranes. To minimize nonspecific binding, the membranes were blocked in 5% bovine serum albumin for 1 hour. The membranes were incubated overnight at 4°C with the following primary antibodies: Iba-1 (1:500; 061-20001, Wako Biologicals), C1q (1:50; ab71940, Abcam), cleaved caspase 3 (1:200; no. 9664S, Cell Signaling Technology), PSD95 (1:200; ab13552, Abcam), synaptophysin (1:500; ab8049, Abcam), or β-actin (1:2500; PA1-183, Thermo Fisher Scientific). After washing, the membranes were incubated with an HRP-conjugated anti-rabbit or anti-mouse antibody (GeneTex Inc., Irvine, CA) for 1 hour. The Western blotting results were visualized with an enhanced chemiluminescence advanced kit. The intensity was analyzed by ImageJ version 1.46 (NIH, Bethesda, MD, USA).

Lee J.S., Lee S.B., Kim D.W., Shin N., Jeong S.J., Yang C.H, & Son C.G. (2021). Social isolation–related depression accelerates ethanol intake via microglia-derived neuroinflammation. Science Advances, 7(45), eabj3400.

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Immunoblotting of β-catenin and Vimentin

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A mouse monoclonal antibody of β-catenin (sc-7963), rabbit polyclonal antibody of vimentin (sc-5565), HRP conjugated goat anti-rabbit IgG (sc-2054), and HRP conjugated goat anti-mouse IgG (sc-2055) were from Santa Cruz Biotech. A rabbit polyclonal antibody of β-actin (PA1183) and Texas Red-X conjugated goat anti-mouse IgG (T6390) were from Thermo Scientific.

Lichtenberger L.M., Fang D., Bick R.J., Poindexter B.J., Phan T., Bergeron A.L., Pradhan S., Dial E.J, & Vijayan K.V. (2016). Unlocking aspirin’s chemopreventive activity: Role of irreversibly inhibiting platelet cyclooxygenase-1. Cancer prevention research (Philadelphia, Pa.), 10(2), 142-152.

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Establishing Primary Dermal Fibroblast Cultures

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We established primary dermal fibroblast cell cultures from epidermal skin punch biopsies from affected individuals and unaffected parents from family 1 and family 2, and RNH1 immunoblotting was performed. We used the following primary antibodies against RNH1 (Novus Biologicals, H0006050-M07; 1:500) and B-actin (ThermoFisher Scientific PA1–183; 1:2000) (Supplemental Data).

Shashi V., Schoch K., Ganetzky R., Kranz P.G., Sondheimer N., Louise Markert M., Cope H., Sadeghpour A., Roehrs P., Arbogast T., Muraresku C., Tyndall A.V., Esser M.J., Woodward K.E., Ping-Yee Au B., Parboosingh J.S., Lamont R.E., Bernier F.P., Wright N.A., Benseler S.M., Parsons S.J., El-Dairi M., Smith E.C., Valdez P., Tennison M., Micheil Innes A, & Davis E.E. (2023). Biallelic variants in ribonuclease inhibitor (RNH1), an inflammasome modulator, are associated with a distinctive subtype of acute, necrotizing encephalopathy. Genetics in medicine : official journal of the American College of Medical Genetics, 25(9), 100897.

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Western Blot Analysis of NLRP3 Inflammasome Proteins

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The protein expressions of NACHT, LRR, and PYD Domains-Containing Protein 3 (NLRP3), apoptosis-associated Speck-like protein containing a CARD (ASC), pro-IL-1β and mature IL-1β, and β-actin in the hippocampal homogenates were evaluated using a western blotting method. The protein concentration of the homogenates was equalized, and the samples were separated by 10% polyacrylamide gel electrophoresis and transferred to polyvinylidene fluoride (PVDF) membranes. To minimize the non-specific binding, the membrane was blocked in 5% bovine serum albumin for 1 h. The membranes were incubated with primary antibodies, such as NLRP3 (1:500, ab214185, Abcam), ASC (1:500, sc-271054, Santa Cruz), pro-IL-1β and mature IL-1β (1:1,000, ab9722, Abcam), or β-actin (1:2500, PA1-183, Thermo-Fisher Scientific), overnight at 4°C. After being washed, the membranes were incubated with an HRP-conjugated anti-rabbit or anti-mouse antibody (GeneTex, Inc., Irvine, CA) for 1 h. The western blotting results were visualized with an enhanced chemiluminescence (ECL) advanced kit. The intensity was analyzed with ImageJ version 1.46 (NIH, Bethesda, MD, USA).

Lee J.S., Kim W.Y., Jeon Y.J., Lee S.B., Lee D.S, & Son C.G. (2019). Antidepressant-Like Activity of Myelophil via Attenuation of Microglial-Mediated Neuroinflammation in Mice Undergoing Unpredictable Chronic Mild Stress. Frontiers in Pharmacology, 10, 683.

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