L polylysine

Manufactured by Merck Group

Sourced in United States

L-polylysine is a linear homopolymer of the amino acid lysine. It is used in various laboratory applications as a coating material to enhance cell adhesion and promote cell growth on surfaces.

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Lab products found in correlation

Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions) Neurobasal medium, by Thermo Fisher Scientific (2 mentions) C57bl 6 female mice, by Merck Group (2 mentions) Alexa fluor 488 goat anti human igg, by Jackson ImmunoResearch (1 mentions) Plan apochromat 63x 1.4 na, by Zeiss (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Vector Laboratories (1 mentions) Vectashield antifade mounting medium, by Vector Laboratories (1 mentions) Dmem f12 medium, by Thermo Fisher Scientific (1 mentions) Mncl2 4h2o, by Merck Group (1 mentions) D hank s solution, by Merck Group (1 mentions) Penicillin streptomycin mixture, by Merck Group (1 mentions) Neurobasal a medium, by Thermo Fisher Scientific (1 mentions) Cell dissociation solution, by Merck Group (1 mentions) Collagenase, by Merck Group (1 mentions) Phrodo green zymosan bioparticles, by Thermo Fisher Scientific (1 mentions) H2dcfda probe, by Thermo Fisher Scientific (1 mentions) H2dcfda, by Thermo Fisher Scientific (1 mentions) Anti tnfα antibody, by Cell Signaling Technology (1 mentions) Trypsin, by Merck Group (1 mentions) Ethylene diamine tetraacetic acid (edta), by Merck Group (1 mentions)

11 protocols using l polylysine

Immunostaining Analysis of Lentiviral-Expressed SARS-CoV-2 Spike

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293T Lenti-X cells (2.5x10⁴/well) were plated onto L-polylysine (Sigma) treated 12-mm cover glasses inserted in 24-well microplates and then transduced with 5 MOI of each LV expressing Spikes used for immunization. At 48 hours post-transduction, the cells were washed and stained with anti-Spike COVA2-15 or COVA1-16 (47 (link)) mAbs followed by AlexaFluor 488 Goat anti-human IgG as a secondary Ab (Jackson ImmunoResearch, 0.4 µg/sample). The coverslips were extensively rinsed, fixed with cold methanol and then placed on the microscope slides using Vectashield antifade mounting medium, containing DAPI (Vector Labs, Burlingame, CA, USA).
Observations by confocal laser scanner mycroscopy (CLSM) were performed using a Zeiss LSM980 apparatus (Zeiss, Oberkochen, Germany), fitted with a Plan-Apochromat 63x/1.4 NA using the appropriate spectral laser lines. Acquisition and processing of images was performed by using Adobe Photoshop CS5 software programs (Adobe Systems, San Jose, CA, USA) and Zen Blue edition 3.3 (Zeiss). Cells which were stained with the secondary antibody were used to set up parameters of acquisition. Several fields (including >200 cells) were evaluated for each labeling condition, and shown are representative results.

Borghi M., Gallinaro A., Pirillo M.F., Canitano A., Michelini Z., De Angelis M.L., Cecchetti S., Tinari A., Falce C., Mariotti S., Capocefalo A., Chiantore M.V., Iacobino A., Di Virgilio A., van Gils M.J., Sanders R.W., Lo Presti A., Nisini R., Negri D, & Cara A. (2023). Different configurations of SARS-CoV-2 spike protein delivered by integrase-defective lentiviral vectors induce persistent functional immune responses, characterized by distinct immunogenicity profiles. Frontiers in Immunology, 14, 1147953.

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Primary Hippocampal Neuron Isolation and Culture

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Primary hippocampal neurons were prepared from E18 Sprague-Dawley (SD) rat embryos and were immediately soaked with 75% ethanol. The hippocampus was dissected from the brain of each rat on ice and stripped the meninges and blood vessels carefully with sterile phosphate-buffered saline (PBS) washing. The tissues were minced and digested with 0.125% trypsin at 37°C for 20 min; digestion was stopped by adding DMEM/F12 medium with 10% fetal bovine serum (FBS) (Gibco, USA). The neurons were centrifuged and suspended to a density of 1 × 10⁶/L in DMEM/F12 (HyClone, USA) with 10% FBS. The different volumes of neuronal suspensions were inoculated in culture flasks and coated with L-poly lysine (Sigma, USA) and cultured in a humidified 5% CO2 atmosphere at 37°C. When the neurons adhered, the medium was changed to neurobasal medium (Gibco, USA). Half of the medium was changed every 3 days. After cultured for 48 h, cytarabine (4 mg/ml) was injected to inhibit the growth of glial cells. Cultured neurons were used for in vitro studies on the 7th day.

Yang L., Wei Y., Luo Y., Yang Q., Li H., Hu C., Yang Y, & Yang J. (2017). Effect of PGE2-EPs pathway on primary cultured rat neuron injury caused by aluminum. Oncotarget, 8(54), 92004-92017.

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Neuronal Cell Culture Reagents

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MnCl₂•4H₂O, L-polylysine, D-Hank’s solution, and penicillin–streptomycin mixture were purchased from Sigma. DMEM-F12 medium was from HyClone. Neurobasal-A medium (without phenol red and serum) and fetal bovine serum (FBS) were purchased from Gibco. Human leukocyte antigen B27 (B27) was obtained from Invitrogen Life Technologies.

Ma S., Qing L., Yang X., Liang G., Zhang L., Li Q., Xiong F., Peng S., Ma Y., Huang X, & Zou Y. (2016). Expression Profiles of Long Noncoding RNAs and Messenger RNAs in Mn-Exposed Hippocampal Neurons of Sprague–Dawley Rats Ascertained by Microarray: Implications for Mn-Induced Neurotoxicity. PLoS ONE, 11(1), e0145856.

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Isolation and Culture of Drosophila Neurons

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Flies were dissected on ice in a sterile Ringers solution and the brains were transferred into ice-cold cell dissociation solution (Sigma, c1544). Brains were incubated with 10mg/ml collagenase (Sigma, c0130), dissolved in cell dissociation solution at 29°C for 30-45 minutes (L3 and pupal brains-30 min, adult brains - 45 min). Next, the brains were centrifuged in 300Xg at 4°C for 6 minutes. The supernatant was discarded and the disintegrated brains were washed with cell dissociation solution. Finally, the cells were resuspended in the culture media which consisted of standard Schneider’s medium (Biological industries) supplemented with 10% heat-inactivated Fetal Bovine Serum (FBS) and antibiotics ([1:100], 100XAntibiotic-Antimyitic, Gibco), mechanically dissociated by pipetting up and down until no aggregates were observed and then kept on ice. The cells were plated on a glass bottom 96-well plate (MatTek, catalog P966-1.5-5-F Case) coated with L-poly-lysine (0.01% solution, Sigma, P4832) and cultured in culturing medium in an incubator at 25°C. When indicated, rapamycin ([100nM], Sigma, R8781) was added. Images were taken using Zeiss LSM 710 confocal microscope, and processed with ImageJ (NIH). Total neurites length or longest neurite per cell were measured using the ImageJ plugin - simple neurite tracer (Longair et al., 2011 (link)).

Marmor-Kollet N, & Schuldiner O. (2015). Contrasting developmental axon regrowth and neurite sprouting of Drosophila mushroom body neurons reveals shared and unique molecular mechanisms. Developmental neurobiology, 76(3), 262-276.

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Quantifying Microglial Phagocytic Activity

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BV2 microglia cells expressing different phenotypes (BV2 WT, BV2^ΔIRF1, and BV2^IRF1-OE) were seeded (25,000 cell/well) into the 96-well plate coated with L-poly-lysine (12.5 ug/mL, 2 h, #P0899, Sigma, Sofia, Bulgaria) and then cultured for 24 h before treatment. The cells were treated with LPS and PBS (as control) for 4h and then incubated with pHrodo™ Green Zymosan Bioparticles™ (100 uL, 0.5 mg/mL, #P35365, Thermo Fisher, Waltham, MA, USA) for 1.5 h. After washing with PBS, the cells were fixed for confocal microscopy and ImageJ analysis to quantify the pixel intensity of Zymosan bioparticles within the cells. Briefly, the fluorescent microscopy images were converted into gray-model images. The pixel number of Zymosan bioparticles was analyzed and determined by the ImageJ software. The pixel intensity quantification was defined as the quotient of the total pixel number divided by the cell number. The averaged pixel intensity per cell represents the phagocytosis activity of microglial cells.

Yang X., Diaz V, & Huang H. (2022). The Role of Interferon Regulatory Factor 1 in Regulating Microglial Activation and Retinal Inflammation. International Journal of Molecular Sciences, 23(23), 14664.

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Orthotopic Murine Model of Bladder Cancer

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Animal experiments were performed according to procedures approved by the Animal Care Committee at the Universitat Autònoma de Barcelona. The orthotopic murine model of BC was developed as previously described^{11 (link),26 (link)}. Briefly, six C57Bl/6 female mice (6–8 weeks old; Charles River Laboratories, France) were anaesthetized and chemical lesions were induced by intravesical instillation of L-poly-lysine (Sigma). Subsequently, 10⁵ MB49 bladder tumor cells were instilled to induce tumors. One day later, each group of mice received intravesically 100 µl of mycobacteria suspensions (corresponding to 2 × 10⁶ colony forming units (CFU) of BCG or 2 × 10⁷ CFU of M. brumae per mouse) or vehicle for control groups. Animals were treated weekly following the schedule shown in Fig. 4 and were sacrificed at day 29 after tumor induction. At the time of sacrifice, blood was collected and bladders and spleens aseptically removed. Bladders from some of the animals of each group were processed for immunohistochemistry (IHQ) analysis, and the other bladders for cytometry studies.Figure 4

Schedule of the animal experiments. Graphical representation of the schedule, in which tumor induction (day 0), mycobacteria treatments (days 1, 8, 15 and 22 after tumor induction) and sacrifice (day 29 after tumor induction) are indicated by arrows.

Noguera-Ortega E., Rabanal R.M., Gómez-Mora E., Cabrera C., Luquin M, & Julián E. (2018). Intravesical Mycobacterium brumae triggers both local and systemic immunotherapeutic responses against bladder cancer in mice. Scientific Reports, 8, 15102.

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Intracellular ROS Detection in Neutrophils

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For the detection of intracellular Reactive Oxidizing Species (ROS), the dichlorodihydrofluorescein diacetate probe (H2DCFDA, Invitrogen) was used. This probe reacts with free radicals in general and emits green fluorescence. For the assays, the 5 × 10⁴ neutrophils were cultured in a 96-well plate treated with 0.001% L-polylysine (Sigma-Aldrich) and incubated at 37 °C/5% CO₂ for 90 min in the presence or absence of stimuli: COVID-19 virus (MOI 9.0); 10% serum from normal or severely infected patients; PMA 100 nM. Quantification of ROS was done by adding 20 μM H2DCFDA probe (Invitrogen) 15 min before the end of incubation. The plates were read at 530 nm emission and 485 nm excitation using a fluorimeter (Spectramax M3).

Arcanjo A., Logullo J., Menezes C.C., de Souza Carvalho Giangiarulo T.C., dos Reis M.C., de Castro G.M., da Silva Fontes Y., Todeschini A.R., Freire-de-Lima L., Decoté-Ricardo D., Ferreira-Pereira A., Freire-de-Lima C.G., Barroso S.P., Takiya C., Conceição-Silva F., Savino W, & Morrot A. (2020). The emerging role of neutrophil extracellular traps in severe acute respiratory syndrome coronavirus 2 (COVID-19). Scientific Reports, 10, 19630.

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Optimized Neuroprotective Protocol

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The Catalpol and Puerarin (purity > 98%) were purchased from Liu bo bai niao Biological Technology Co., Ltd (Hebei, China). The lyophilized Powder of Catalpol and Puerarin was prepared in our Lab. Fetal Bovine Serum (FBS), DMEM/F12, and B27 were purchased from GIBCO (Gaithersburg, MD, USA). MTT, trypsin, L-glutamine, EDTA, gelatin, sodium fluorescein (SF), Collagenase type II、Bull Serum Albumin (BSA) and L-polylysine were purchased from Sigma-Aldrich (St Louis, MO, USA). Except that the anti-TNF-α antibody was from Cell Signaling Technology, all the antibodies used in this research were purchased from Proteintech Group, Inc. (Chicago, IL, USA) and kits from YUANYE-BIO (Shanghai, China)

XUE Q., LIU Y., HE R., YANG S., TONG J., LI X., CHEN Y, & XU X. (2016). Lyophilized Powder of Catalpol and Puerarin Protects Neurovascular Unit from Stroke. International Journal of Biological Sciences, 12(4), 367-380.

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Primary Hippocampal Neuron Isolation

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Primary hippocampal neurons were prepared from E18 rat embryos and were immediately soaked with 75% ethanol. The hippocampus was isolated from the brain of each rat and the tissues were minced and digested with 0.125% trypsin at 37 °C for 20 min; digestion was stopped with the addition of 10% fetal bovine serum (FBS) (Gibco, USA). The neurons were centrifuged and suspended to a density of 1 × 10⁶/L in DMEM (HyClone, USA) with 10% FBS in it. The different volumes of neuronal suspensions were inoculated in culture flasks and coated with L-poly lysine (Sigma, USA) and cultured in a humidified 5% CO₂ atmosphere at 37 °C. When the neurons adhered, the medium was changed to neurobasal medium (Gibco, USA).

Ma J., Yang Q., Wei Y., Yang Y., Ji C., Hu X., Mai S., Kuang S., Tian X., Luo Y., Liang G, & Yang J. (2016). Effect of the PGD2-DP signaling pathway on primary cultured rat hippocampal neuron injury caused by aluminum overload. Scientific Reports, 6, 24646.

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Visualizing Protein Localization in 293T Cells

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293T Lenti-X cells (2 × 10⁴/well) were seeded in 24-well cluster plates onto 12-mm cover glasses previously treated with l-polylysine (Sigma) and transfected with plasmids using the Profection Mammalian Transfection System (Promega). Twenty-four hours post-transfection, cells were washed and fixed with 4% paraformaldehyde (PFA) for 30 min at +4°C, and the coverslips were mounted with Vectashield antifade mounting medium-containing DAPI (Vector Labs, Burlingame, CA, USA) on the microscope slides. CLSM observations were performed on a Leica TCS SP2 AOBS apparatus (Leica Microsystems, Wetzlar, Germany) using excitation spectral laser lines at 488 and 546 nm (for GFP) or at 586 and 610 nm (for mCherry), and using the confocal software (Leica, Wetzlar, Germany) and Photoshop CS5 (Adobe Systems, San Jose, CA, USA). Signals from different fluorescent probes were taken in sequential scanning mode, several fields were analyzed for each labeling condition, and representative results are shown. Images represented a single central optical section taken in the center of each cell nucleus.

Gallinaro A., Borghi M., Pirillo M.F., Cecchetti S., Bona R., Canitano A., Michelini Z., Di Virgilio A., Olvera A., Brander C., Negri D, & Cara A. (2020). Development and Preclinical Evaluation of an Integrase Defective Lentiviral Vector Vaccine Expressing the HIVACAT T Cell Immunogen in Mice. Molecular Therapy. Methods & Clinical Development, 17, 418-428.

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