Mammalian protein extraction reagent m per buffer

Manufactured by Thermo Fisher Scientific

Sourced in United States

Mammalian Protein Extraction Reagent (M-PER) buffer is a solution designed for the extraction of proteins from mammalian cells and tissues. It is a non-denaturing, mild detergent-based buffer that helps solubilize cellular proteins while maintaining their native structure and function.

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Lab products found in correlation

Phosstop, by Roche (2 mentions) Halt protease inhibitor cocktail, by Thermo Fisher Scientific (1 mentions) Pierce bicinchoninic acid bca protein assay, by Thermo Fisher Scientific (1 mentions) Ficoll pm70, by Merck Group (1 mentions) Phosphate buffered saline (pbs), by Merck Group (1 mentions) 2 3 cgamp elisa kit, by Cayman Chemical (1 mentions) Anti gfp, by Merck Group (1 mentions) Ib23002, by Thermo Fisher Scientific (1 mentions) Ib21001, by Thermo Fisher Scientific (1 mentions) Anti myc, by Santa Cruz Biotechnology (1 mentions) Turbofect, by Thermo Fisher Scientific (1 mentions) Pvdf membrane, by Merck Group (1 mentions) Bca protein assay kit, by Thermo Fisher Scientific (1 mentions) Protease inhibitor cocktail, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

M-PER (386 citations) Mammalian Protein Extraction Reagent (211 citations) MES buffer (151 citations) M-PER buffer (125 citations) Protein extraction reagent (96 citations) M-PER reagent (72 citations) Extraction buffer (35 citations) M-PER extraction buffer (12 citations) Protein extraction buffer (11 citations) M-PER Mammalian Extraction Buffer (10 citations)

6 protocols using mammalian protein extraction reagent m per buffer

Evaluating Inhibitory Effects on Oxidative Stress

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After 24 h incubation, culture media were removed and cultures were exposed to the inhibitors MI432 and MI460 for 4 and 24 h at 10, 25, and 50 µM concentrations. The structures (Figure 2) and inhibitory constant (Ki) values (Table 1) of the applied MI432 and MI460 inhibitors were already published by Hammami et al. [24 (link)]. All 3-APhA inhibitors were kindly provided by the Institute of Pharmaceutical Chemistry, Faculty of Pharmacy, Philipps University, Marburg, Germany.
Culture media of 6-well plates were collected directly after the incubation time to assess extracellular H₂O₂, IL-6, IL-8, and MDA concentrations, while cells were lysed in Mammalian Protein Extraction Reagent (M-PER) buffer supplemented with 1% Halt Protease Inhibitor Cocktail and 1% ethylene diamine tetraacetic acid (EDTA) (Thermo Fisher Scientific, Waltham, MA, USA). To standardize the results, total protein concentrations of cell lysates were measured with Pierce Bicinchoninic Acid (BCA) Protein Assay (Thermo Fisher Scientific, Waltham, MA, USA), using BSA as a standard, following the manufacturer’s description. All samples were stored at −80 °C until further processing.

Barna R.F., Mackei M., Pászti-Gere E., Neogrády Z., Jerzsele Á, & Mátis G. (2021). The Effects of Matriptase Inhibition on the Inflammatory and Redox Homeostasis of Chicken Hepatic Cell Culture Models. Biomedicines, 9(5), 450.

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Optimizing Protein Extraction Buffer

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Buffers for in vitro experiments consisted of a mixture of Ficoll PM 70 (Sigma-Aldrich, St. Louis, MO) between 0 and 300 mg/mL and Mammalian-Protein Extraction Reagent (M-PER™) buffer (ThermoFisher Scientific, Waltham, MA) between 0 and 90% v/v. Stock Ficoll solutions of ≈400 mg/mL were prepared by stirring Ficoll into sodium phosphate buffer or M-PER™ buffer and left in a water bath at 55°C overnight. The stock was allowed to come to room temperature and dilutions were prepared for in vitro measurements. To ensure complete solubilization of Ficoll, the final concentration of Ficoll and M-PER™ buffer was sonicated for 10 min immediately prior to the addition of protein and stability measurement.

Knab E, & Davis C.M. (2023). Chemical interactions modulate λ_6-85 stability in cells. Protein science : a publication of the Protein Society, 32(7).

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Quantifying cGAMP in Cell Lines

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For cGAMP quantification, T98G^Empty, T98G^cGAS, THP‐1^CTRL, and THP‐1^cGAS−/− were seeded 18 h before dsDNA transfection. One hour before dsDNA transfection (1 μg/ml), cells were pretreated with 2 μM NU7441 (#3712, Biotechne/Tocris) in OptiMEM. Cells were harvested 6 h post transfection, counted, washed in phosphate‐buffered saline (PBS) (Sigma), pelleted, and frozen at −80°C until extraction. cGAMP extraction was performed using the commercially available Mammalian Protein Extraction Reagent (M‐PER) buffer (Thermo Fisher), accordingly to the manufacture protocol. The recovered supernatants were used for cGAMP measurement, following adequate sample dilution. cGAMP enzyme‐linked immunosorbent assay (ELISA) was performed according to the manufacturer's protocol using the Cayman Chemical 2′3′‐cGAMP ELISA Kit (CAY501700).

Taffoni C., Marines J., Chamma H., Guha S., Saccas M., Bouzid A., Valadao A.C., Maghe C., Jardine J., Park M.K., Polak K., De Martino M., Vanpouille‐Box C., Del Rio M., Gongora C., Gavard J., Bidère N., Song M.S., Pineau D., Hugnot J., Kissa K., Fontenille L., Blanchet F.P., Vila I.K, & Laguette N. (2022). DNA damage repair kinase DNA‐PK and cGAS synergize to induce cancer‐related inflammation in glioblastoma. The EMBO Journal, 42(7), e111961.

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Protein Extraction from Drosophila Heads

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Briefly, 40 to 50 heads were collected from flies and ground in M-PER mammalian protein extraction reagent buffer (ThermoFisher Scientific, Waltham, MA, USA, #78501, country) supplemented with protease inhibitor (Roche, Basel, Switzerland #11697498001) and PhosStop (Roche #4906845001) and placed on ice for 30 min. They were centrifuged at maximum speed for 15 min, and the supernatant was collected for Western blot. Protein was extracted from fly head homogenates, and equal amounts of protein from the various genotypes were resolved by SDS-PAGE and transferred onto nitrocellulose membrane using an Iblot 2 transfer device (ThermoFisher Scientific #IB21001 and #IB23002). The following antibodies are used for probing the blot for LRRK2: anti-GFP (Sigma Aldrich, St. Louis, MO, USA #G1544), anti-myc (Santa Cruz Biotechnology, Santa Cruz, CA, USA #sc-40)

Toh J., Chua L.L., Ho P., Sandanaraj E., Tang C., Wang H, & Tan E.K. (2021). Identification of Targets from LRRK2 Rescue Phenotypes. Cells, 10(1), 76.

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GFP-Protein Immunoprecipitation from HEK293T

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HEK293T cells were co-transfected with the plasmids using Turbofect (Thermo Scientific). Cells were collected 24h after transfection for western blot. Transfected HEK293T cells were washed with PBS and lysed in M-PER mammalian protein extraction reagent buffer (Thermo Scientific) supplemented with protease inhibitor and Phos Stop (Roche). The lysates are then incubated with anti GFP conjugated beads (Chemotek) O/N at 4ºC. The precipitates were then washed 5 times using NP40 buffer (50mM Tris pH7.4, 150mM NaCl and 1% NP40) and resuspended in 4X SDS loading buffer.

Chua L.L., Ho P., Toh J, & Tan E.K. (2020). Chetomin rescues pathogenic phenotype of LRRK2 mutation in drosophila. Aging (Albany NY), 12(18), 18561-18570.

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Immunoblotting Analysis of YAP1 Signaling

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Tissue samples were collected and lysed with M-PER mammalian protein extraction reagent buffer (Thermo Fisher Scientific, San Jose, CA, USA) containing protease inhibitor cocktail (Thermo Fisher Scientific, Rockford, Illinois, USA) in ice-cold temperature. After 30 min of lysis, the proteins were extracted by centrifugation at 13,000 g for 15 min. The protein concentrations were determined using the BCA protein assay kit (Thermo Fisher Scientific, San Jose, CA, USA). Equal amounts of protein (40 μg) from each sample were separated with 10% sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS–PAGE) and electrophoretically transferred to PVDF membranes (Millipore, Boston, MA, USA) for immunoblotting. After being blocked with 5% nonfat milk in TBST for 1 h at room temperature, the blots were incubated overnight at 4°C with primary antibodies against human YAP1 (1 : 1000, ET1608-30), p-YAP1 (1 : 1000, ET1611-69), and GAPDH (1 : 5000, EM1101). All antibodies were obtained from Huabio Technology (Hangzhou, China). The immunoreactive protein blots were visualized using SuperSignal reagents (ECL; Millipore, Billerica, MA, USA), and the results were analyzed with ImageJ software.

Yao W., Ma A., Zhang Z, & Zhu L. (2022). Intermittent Hydrostatic Pressure Promotes Cartilage Repair in an Inflammatory Environment through Hippo-YAP Signaling In Vitro and In Vivo. BioMed Research International, 2022, 3215461.

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