P1379

Manufactured by Merck Group

Sourced in United States, Germany, United Kingdom

The P1379 is a laboratory equipment product manufactured by Merck Group. It serves as a general-purpose tool for various laboratory applications. The core function of this product is to provide a reliable and consistent platform for conducting experiments and analyses. Additional details about its intended use or specific applications are not available.

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Lab products found in correlation

A7906, by Merck Group (5 mentions) T8787, by Merck Group (4 mentions) Lsm 880, by Zeiss (4 mentions) Skimmed milk, by Merck Group (3 mentions) Clarity western ecl substrate, by Bio-Rad (3 mentions) D9542, by Merck Group (3 mentions) Paraformaldehyde (pfa), by Merck Group (3 mentions) Axio observer z1, by Zeiss (3 mentions) Bca kit, by Thermo Fisher Scientific (3 mentions) Chloramphenicol, by Merck Group (2 mentions) Kanamycin, by Merck Group (2 mentions) Ampicillin, by Merck Group (2 mentions) Triton x 100, by Merck Group (2 mentions) Sc 47778, by Santa Cruz Biotechnology (2 mentions) Ab49900, by Abcam (2 mentions) Ab6721, by Abcam (2 mentions) Immobilon p, by Merck Group (2 mentions) H 1200, by Vector Laboratories (2 mentions) Trans blot turbo transfer system, by Bio-Rad (2 mentions) Chemidoc mp system, by Bio-Rad (2 mentions)

Close spelling variants of this product

P1379-500ML (3 citations) P1379-25ML (2 citations) P1379-100ML (2 citations)

75 protocols using p1379

In situ Hybridization of Retinal Genes

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For in toto hybridization, we collected retinas after fixation as described above and dehydrated them with increasing concentrations of methanol (25%, 50%, 75% and 100%) in 0.1% PBS-Tween 20 (Sigma, P1379). For in situ hybridization on sections, we directly embedded and froze the eyes in Optimal Cutting Temperature compound without fixation. 20-μm sections were cut with a cryostat.
Antisense riboprobes were labeled with digoxigenin-11D-UTP (Roche) as described previously^{46 (link)} by in vitro transcription of mouse cDNAs encoding Slit1 (ref. 47 (link)), Slit2 (ref. 48 (link)), Slit3 (nucleotides 2,270–4,642) or Plxnd1 (ref. 49 (link)). A mouse Slit2 cDNA specific for exons 8–9 was amplified by PCR and cloned into pBluescript. Whole-mount retinas and retinal sections were hybridized as described previously^{48 (link)}.
In situ hybridization images were obtained with a DM6000 microscope (Leica) and CoolSNAP CCD camera (Roper).

Rama N., Dubrac A., Mathivet T., Chárthaigh R.A., Genet G., Cristofaro B., Pibouin-Fragner L., Ma L., Eichmann A, & Chédotal A. (2015). Slit2 signaling through Robo1 and Robo2 is required for retinal neovascularization. Nature medicine, 21(5), 483-491.

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E. coli Strain 39 Microfluidic Cultivation

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Experiments with bacteria used a previously generated E. coli strain 39 (link) . Luria-Bertani (LB) medium (113002065, MP Biomedicals) supplemented with 50 µg/mL kanamycin (K4000, Sigma-Aldrich), 100 µg/mL ampicillin (A9518, Sigma-Aldrich) and 25 µg/mL chloramphenicol (C0378, Sigma-Aldrich) was used for all bacterial cell culture and microfluidics experiments.
For microfluidic experiments, a single colony was used to seed 5 mL of LB media with antibiotics and grown overnight (approximately 16 hours) at 37°C with shaking at 200 rpm. 300 µL of the overnight culture was used to seed 300 mL of fresh LB medium with antibiotics. This culture was grown to an optical density at 600 nm of 0.3. The culture was then centrifuged at 2200 × g for 15 min and resuspended in 1.5 mL of fresh LB medium supplemented with 0.075% Tween-20 (P1379, Sigma-Aldrich) and antibiotics before loading into the microfluidic device.

Pedone E., de Cesare I., Zamora-Chimal C.G., Haener D., Postiglione L., La Regina A., Shannon B., Savery N.J., Grierson C.S., di Bernardo M., Gorochowski T.E, & Marucci L. (2021). Cheetah: A Computational Toolkit for Cybergenetic Control. ACS synthetic biology, 10(5).

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Comparative Microscopy Imaging of Stained Cells

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To compare image quality acquired with our microscope and a conventional microscope, human monocytic (THP-1, kindly donated by Dr. Vianney Ortiz) cells were permeabilized with Tween-20 (P1379, Sigma Aldrich) 0.05% in Phosphate-Buffered Saline (PBS) 1X for 15 min, centrifuged at 1,200 rpm for 5 min, and washed with PBS 1X. Next, cells were incubated for 20 min either with 4′,6-diamidino-2-phenylindole (DAPI, 2.9 μM, 62247, Thermo Fisher Scientific) or Calcein-AM (20 μM, C1359, Sigma Aldrich), or for 10 min with Ethidium Homodimer (16 μM, EthD-1, E1903 Sigma Aldrich). The cells were washed again with PBS 1X and centrifuged at 1,200 rpm for 5 min. Then, 20 μL of the cellular suspensions were spread over three different clean coverslips and allowed to dry. Finally, 10 μL of glycerol were added and a second coverslip was placed on the top of the samples. For Calcein-AM staining, cells were not permeabilized, and PBS 1X was used instead of glycerol. Finally, coverslips were sealed with enamel.

Tristan-Landin S.B., Gonzalez-Suarez A.M., Jimenez-Valdes R.J, & Garcia-Cordero J.L. (2019). Facile assembly of an affordable miniature multicolor fluorescence microscope made of 3D-printed parts enables detection of single cells. PLoS ONE, 14(10), e0215114.

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E. coli Strain 39 Microfluidic Cultivation

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Pedone E., Cesare I.d., Zamora-Chimal C.G., Haener D., Postiglione L., Regina A.L., Shannon B., Savery N.J., Grierson C., Bernardo M.d., Gorochowski T.E., & Marucci L. (2020). Cheetah: a computational toolkit for cybergenetic control.

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Immunoprecipitation and Western Blot Analysis Protocol

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HEK293T cells seeded in 10-cm-diameter dishes were cotransfected with the indicated expression vectors (5 μg each) using Lipofectamine 3000. At 48 h post-transfection, the cells were collected and lysed in 500 μl lysis buffer (P0013; Beyotime) on ice for 1 h. The lysates were harvested by centrifugation at 12,000 g, 4°C for 10 min. Protein beads (15920010; Life Technologies) were incubated with the indicated antibodies at room temperature for 1 h, added to the cell lysates, and incubated at 4°C overnight. After four washes with lysis buffer, the precipitated proteins were eluted from the beads and used for immunoblot analysis.
For western blotting, the isolated cellular protein was denatured at 95°C for 10 min. The cell lysates were resolved by 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred to nitrocellulose membranes. The membranes were blocked using 5% bovine serum albumin (BSA) dissolved in Tris-buffered saline containing 0.2% Tween-20 (TBST; P1379; Sigma) at room temperature for 2 h. Then, the membranes were incubated with primary antibodies at 4°C overnight. After three washes with TBST, the membranes were incubated with secondary antibodies at room temperature for 1 h. After another three washes with TBST, the proteins on the membrane were visualized using an enhanced chemiluminescence reagent (WBKLS0500; Millipore).

Yao Y.L., Luo Y., Wang Q., Geng R., Chen Y., Liu M.Q., Li B., Chen J., Wu C.G., Jia J.K., Luo J.Y., He Y.T., Jiang T.T., Zhu Y., Hu B., Zhou P, & Shi Z.L. (2023). Identification of TMEM53 as a novel SADS-CoV restriction factor that targets viral RNA-dependent RNA polymerase. Emerging Microbes & Infections, 12(2), 2249120.

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Detection of aMPV/C Infection in Vero Cells

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Vero cells with 90% confluence were infected with aMPV/C (MOI = 0.5) in 24-well culture plates. The cells were fixed with precooled and permeabilized using 4% paraformaldehyde (Sigma-Aldrich, 16005), 0.1% Triton X-100 (Sigma-Aldrich, T8787) and in 2% BSA (Beyotime, ST023) in PBS at different indicated time points, and anti-N monoclonal antibody and anti-MAVS rabbit polyclonal antibody were co-incubated with the cells for 2 h at 37 °C. After 3 washes with PBS-Tween-20 (PBST containing 0.05% Tween-20 [Sigma, P1379]), the cells were co-incubated with secondary fluorescein isothiocyanate (FITC)-conjugated antirabbit and Tetramethylrhodamine-6-isothiocyanate (TRITC)-conjugated antimouse antibodies for 2 h at 37 °C. Finally, the cells were washed with PBST and directly observed under an Olympus IX73 immunofluorescence microscope.

Hou L., Hu X., Guo J., Quan R., Wei L., Wang J., Song J, & Liu J. (2021). Avian Metapneumovirus Subgroup C Induces Mitochondrial Antiviral Signaling Protein Degradation through the Ubiquitin-Proteasome Pathway. Viruses, 13(10), 1990.

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Western Blot Analysis of Exosomal Proteins

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Eluates and cell or exosome lysates were separated by SDS-PAGE electrophoresis in gradient 4–12% NuPAGE gels (Invitrogen, NP0321) and transferred onto PVDF membrane (Bio-Rad, 162-0264) using Western blot. Membranes were blocked for 1 h in Blotting-Grade Blocker (Bio-Rad, 165-3301) and subsequently incubated with primary antibodies for 1 h at room temperature. The TBS (tris-buffered saline) buffer supplemented with 0.1% Tween 20 (Sigma-Aldrich, P1379) was used in all steps. Rabbit anti-UCH-L5 (Abcam, ab133508), rabbit anti-USP10 (Cell Signaling Technology, 8501), rabbit anti-USP25 antibody (Abcam, ab187156), and mouse anti-tubulin antibodies (Abcam, ab59680), respectively, were used for UCH-L5, USP10, USP25, and tubulin detection. Rabbit anti-CD9 (Cell Signaling Technology, 13174) was used for detection of the CD9 exosome marker.
After incubation with appropriate horseradish peroxidase-labeled secondary antibodies and washing membranes in TBS supplemented with 0.1% Tween 20, proteins were detected using enhanced chemiluminescence reagents (GE Healthcare, RPN2209 or Thermo Fisher Scientific, 34094). Images were visualized on medical X-Ray blue film (AGFA NV, XDAOG).

Vozandychova V., Rehulka P., Hercik K., Spidlova P., Pavlik P., Hanus J., Hadravova R, & Stulik J. (2023). Modified activities of macrophages’ deubiquitinating enzymes after Francisella infection. Frontiers in Immunology, 14, 1252827.

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Quantitative Western Blot Analysis of Apoptosis Regulators

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Briefly, MDA‐MB‐453, MCF‐7, and MDA‐MB‐231 cell lines or neonatal cardiomyocytes were homogenized in ice‐cold lysis buffer. After centrifugation at 5000g for 20 min, protein content of the supernatant was quantified using the Bradford protein assay. Samples were diluted, boiled with sample loading dye, and 100 μg were loaded in SDS‐PAGE (4561033EDU; Bio‐Rad). After blotting, membranes were blocked in 5% skim milk (70166; Sigma‐Aldrich) in PBS containing 0.1% Tween‐20 (P1379; Sigma‐Aldrich). Membranes were incubated with antisera directed against cytochrome C (1:1000; #11940, Cell Signaling Technology, USA), Endo G (1:1000; #4969, Cell Signaling Technology), or AIF (1:1000; ab1998, Abcam, USA), then with secondary antibodies (mouse‐specific HRP‐conjugated antibody or rabbit‐specific HRP‐conjugated antibody). Bands were visualized using ECL (32106, Thermo Scientific) detection kit and quantified by densitometry. Blots were stripped and re‐exposed to detect β‐actin (1:1000; sc‐47778, Santacruz Biotechnology, USA) as housekeeping protein.

Kim H., Ahn Y., Moon C.M., Kang J.L., Woo M, & Kim M. (2022). Lethal effects of mitochondria via microfluidics. Bioengineering & Translational Medicine, 8(3), e10461.

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Western Blot Analysis of SARS-CoV-2 Spike Protein

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Cells were washed gently with 1× warm PBS (162528, MP Biomedicals), lysed using 1× Laemmli buffer (1610747, BIO-RAD), and heated at 95°C before loading on to a 10% SDS-PAGE gel. Separated proteins were transferred onto a PVDF membrane (IPVH00010, Immobilon-P; Merck) and incubated for 2 hr with blocking buffer containing 5% Skimmed milk (70166, Sigma-Aldrich) in PBST (1× PBS containing 0.05% Tween 20 (P1379, Sigma-Aldrich)) for 2 hr at RT (room temperature). The blots were then probed with SARS-CoV-2 spike antibody (NR-52947, BEI Resources, NIAID, NIH) in blocking buffer for 12 hr at 4°C, followed by secondary Goat Anti-Rabbit IgG antibody (ab6721, Abcam, RRID:AB_955447) incubation for 2 hr. Proteins were detected using Clarity Western ECL Substrate (1705061, BIO-RAD). Actin was labeled using antibody against beta-actin [AC-15] (HRP) (ab49900, Abcam, RRID: AB_867494). Relative intensity of bands was quantified using imagej/Fiji.

Afsar M., Narayan R., Akhtar M.N., Das D., Rahil H., Nagaraj S.K., Eswarappa S.M., Tripathi S, & Hussain T. (2022). Drug targeting Nsp1-ribosomal complex shows antiviral activity against SARS-CoV-2. eLife, 11, e74877.

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Immunofluorescent Analysis of hPSC-CMs

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The hPSC-CMs on coverslips were washed twice with cold PBS, and fixed with 4% paraformaldehyde on ice for 10 min. Fixed hPSC-CMs were washed 3 times with PBS, followed by permeabilization with PBS + 0.3% Triton-X100 (T9284, Sigma-Aldrich (St. Louis, MO, USA)) on ice for 5 min. Samples were blocked for 1 h at room temperature with PBS/Tween (0.1%; P1379, Sigma-Aldrich (St. Louis, MO, USA)) containing 3% BSA (11930, Serva (Heidelberg, Germany)) and 2% donkey serum (D9663, Sigma (St. Louis, MO, USA)). The hPSC-CMs were subsequently incubated with polyclonal anti-cardiac troponin T IgG (1:100; ab45932, Abcam(Cambridge, UK)), Total OXPHOS Rodent WB Antibody Cocktail (1:400; ab110413, Abcam (Cambridge, UK)) and monoclonal anti-Ki-67 (1:200; MA5-14520, Thermo Fisher Scientific (Waltham, MA, USA)) diluted in the blocking mix for 1 h. After washing, the cells were incubated with Alexa Fluor 488 donkey-anti-mouse IgG (1:1000; A21202, Thermo Fisher Scientific (Waltham, MA, USA)) or Alexa Fluor 555 donkey-anti-rabbit IgG (1:1000; A31572, Thermo Fisher Scientific (Waltham, MA, USA)). Coverslips were mounted with Vectashield mounting medium containing DAPI (H-1200, Vector labs (Burlingame, CA, USA)) and images were obtained with a Leica AF-6000 microscope (Leica (Wetzlar, Germany)).

Bomer N., Pavez-Giani M.G., Deiman F.E., Linders A.N., Hoes M.F., Baierl C.L., Oberdorf-Maass S.U., de Boer R.A., Silljé H.H., Berezikov E., Simonides W.S., Westenbrink B.D, & van der Meer P. (2021). Selenoprotein DIO2 Is a Regulator of Mitochondrial Function, Morphology and UPRmt in Human Cardiomyocytes. International Journal of Molecular Sciences, 22(21), 11906.

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