SHED cells were seeded into 75 cm2 flasks at a density of 2 × 106 cells per flask in a growth culture medium and allowed to attach for 24 h. Then, the cells were treated with PEG-AuNR or PEG-AuNS suspensions (0.5 nM) in the tissue culture medium containing FBS and serum-free medium (SFM) was immediately applied to the cells for 6 h. The media was discarded after 6 h of incubation, and the cells were washed with PBS. The cells were fixed with 10% PFA for 10 min. Then, the fixed cells were washed thrice with the washing buffer. After that, 4′,6-diamidino-2-phenylindole (DAPI) stain (Thermofisher, Waltham, MA, USA) was added to the cells and incubated for 5 min, followed by a washing step with PBS. Finally, coverslips were transferred onto glass slides loaded with one drop of mounting media (DAKO, Glostrup, Denmark). Confocal images were acquired via a laser scanning microscope 780 (Zeiss, Oberkochen, Germany). The objective used for acquiring the images was a Plan-Apochromat 63X/1.4 Oil DIC M27. The cells were imaged at excitation/emission wavelengths of 532 nm/750 nm for gold and 360 nm/460 nm for DAPI.
4 6 diamidino 2 phenylindole dapi stain
Manufactured by Thermo Fisher Scientific
Sourced in United States
4,6-diamidino-2 phenylindole (DAPI) is a fluorescent stain that binds strongly to adenine-thymine (A-T) rich regions in DNA. It is commonly used in various biological applications to visualize and identify cell nuclei.
Automatically generated - may contain errors
Lab products found in correlation
Naio4, by Merck Group (2 mentions)
Ir 780 iodide, by Merck Group (2 mentions)
Poly l lysine coated glass slides, by Avantor (2 mentions)
1 1 dioctadecyl 3 3 3 3 tetramethylindodicarbocyanine 4 chlorobenzenesulfonate salt did, by Thermo Fisher Scientific (2 mentions)
Aldehyde quantification assay kit fluorometric, by Abcam (2 mentions)
Glycerol, by Merck Group (2 mentions)
Bovine serum albumin (bsa), by Merck Group (2 mentions)
Mounting media, by Agilent Technologies (1 mentions)
Laser scanning microscope 780, by Zeiss (1 mentions)
Crystal violet, by HiMedia (1 mentions)
Phosphate buffered saline (pbs), by Thermo Fisher Scientific (1 mentions)
Mueller hinton broth (mhb), by HiMedia (1 mentions)
Gram staining kit, by HiMedia (1 mentions)
Isopropanol, by Merck Group (1 mentions)
Methanol, by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Sodium chloride (nacl), by Merck Group (1 mentions)
Absolute ethanol, by Merck Group (1 mentions)
Nutrient agar, by HiMedia (1 mentions)
3 4 5 dimethylthiazol 2 yl 2 5 diphenyltetrazolium bromide, by Merck Group (1 mentions)
11 protocols using 4 6 diamidino 2 phenylindole dapi stain
1
Uptake of Gold Nanoparticles in SHED Cells
2
Microbial Characterization and Cell Viability
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Mueller Hinton Broth (MHB), Nutrient Agar (NA), Gram Staining Kit, and Crystal Violet (cell culture tested) used in the study were supplied by HiMedia. Dimethyl sulfoxide (DMSO), Methanol, Glycerol [Aqueous] acetic acid, Sodium Chloride (NaCl), Isopropanol, Absolute Ethanol, Resazurin, and 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyl tetrazolium bromide (MTT) was purchased from Merck (India). 4′,6-diamidino-2-phenylindole (DAPI) Stain and Propidium Iodide counter stain (PI) were purchased from Thermo Fischer Scientific. Foetal Bovine Serum (FBS), Bovine Serum Albumin (BSA), Dulbecco’s Modified Eagle Medium (DMEM), Trypsin, phosphate-buffered saline (PBS) were obtained from Gibco (Thermo Fischer Scientific). All chemicals used were of analytical research grade.
3
Quantifying Apoptosis in Tissue Samples
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Apoptotic cells in the normal tissue and tumor tissue sections were evaluated using an apoptosis detection kit (Takara Biomedicals, Tokyo, Japan) and expressed as the number of terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL)-positive cells.
22 (link)
The number of TUNEL-positive cells in five visual fields was counted under a fluorescence microscope using Image J software. Tissues were treated with 4',6-diamidino-2-phenylindole (DAPI) stain (Thermo Fisher Scientific; 0.5 μg/mL in TBS) for 3 minutes and washed three times with 2 mL TBS and dried, following which a non-fluorescent encapsulant was added (Fluoromount Aqueous Mounting Medium; Empire Genomics LLC, Buffalo, New York, United States).
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Apoptotic cells in the normal tissue and tumor tissue sections were evaluated using an apoptosis detection kit (Takara Biomedicals, Tokyo, Japan) and expressed as the number of terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL)-positive cells.
22 (link)
The number of TUNEL-positive cells in five visual fields was counted under a fluorescence microscope using Image J software. Tissues were treated with 4',6-diamidino-2-phenylindole (DAPI) stain (Thermo Fisher Scientific; 0.5 μg/mL in TBS) for 3 minutes and washed three times with 2 mL TBS and dried, following which a non-fluorescent encapsulant was added (Fluoromount Aqueous Mounting Medium; Empire Genomics LLC, Buffalo, New York, United States).
4
F-actin Visualization in AsPC-1 Cells
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For F-actin staining, AsPC-1 cells were cultured in CytoSoft® 6-well plates or on fabricated polyacrylamide gels for 12 h. After 12 h, the cells were fixed with 4% paraformaldehyde (PFA) for 15 min, rinsed three times with 1× PBS, permeabilized with 0.1% Triton-X 100 (Thermo Fisher Scientific, Waltman, Massachusetts, USA) for 10 min, and then rinsed again 3 times with 1× PBS. Alexa Fluor® 555 phalloidin Ex/Em 555/565 nm (Abcam, Cambridge, UK) was used according to the manufacturer's instructions. Nuclei were visualized using 4′,6-diamidino-2-phenylindole (DAPI) stain in a 1 μg ml−1 solution (Thermo Fisher Scientific, Waltman, Massachusetts, USA). Images were obtained using an inverted fluorescent microscope (model IX73, Olympus Corporation, Japan) and a confocal microscope (Nikon's A1 MP multiphoton confocal microscope equipped with a 639-nm diode).
5
Histological Analysis of Islet Grafts
6
Immunofluorescent Staining of Cytoskeleton
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NIH3T3 cells in six‐well plates were washed twice with cold PBS and then fixed with methanol/acetone (v : v = 1 : 1) followed by washing with PBS. After blocking in 1% BSA for 1 h at room temperature, cells were incubated with a mouse anti‐tubulin antibody (Abcam, Cambridge, MA, USA) for 1 h. Following washing with PBS, cells were incubated with Alexa Fluor 594‐conjugated secondary antibody followed by 4′,6‐diamidino‐2‐phenylindole (DAPI) stain (Life Technologies, Foster City, CA, USA). Finally, cells were imaged using a laser scanning fluorescence microscope (Olympus, Tokyo, Japan) 15 .
7
Immunocytochemistry of Lgr5 and C. parvum
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Cell cultures were fixed [0.1 mol/L, piperazine‐1,4‐bis(2‐ethanesulphonic acid) (Sigma‐Aldrich), pH 6.95, 1 mmol/L ethylene glycol‐bis (2‐oiminoether)‐ N,N,N′,N′‐tetraacetic acid] (Sigma‐Aldrich), 3 mmol/L magnesium sulphate (Sigma‐Aldrich) and 2% paraformaldehyde] at room temperature for 35 min and then permeabilized with 0.1% (v/v) Triton X‐ 100 in PBS. Fixed cultures were then incubated with primary antibodies and secondary antibodies, according to the standard immunochemical approach (Chen et al. 2007 ). Labelled cultures were then mounted using ProLong Gold Antifade Reagent with 4, 6‐diamidino‐2‐phenylindole (DAPI) stain (Invitrogen), followed by fluorescence microscopy. Anti‐Lgr5 (5 ng/mL, Upstate) and anti‐C. parvum (Zhou et al. 2012 ) were used. For localization of actin, fluorescein‐phalloidin (Sigma‐Aldrich) was incubated for 30 min at room temperature after incubation with the primary antibodies.
8
Characterizing Sunscreen Composition and Effects
9
Characterizing Sunscreen Composition and Effects
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IR-780 iodide, glycerol, NaIO4 and Bovine serum albumin (BSA) were obtained from the Sigma-Aldrich. The 1,1'-Dioctadecyl-3,3,3',3'-Tetramethylindodicarbocyanine,4 Chlorobenzenesulfonate Salt (DiD) and 4,6-diamidino-2 phenylindole (DAPI) stain were ordered from Invitrogen. Aldehyde Quantification Assay Kit (Fluorometric) was from Abcam. Poly -L -lysine coated glass slides were obtained from VWR International Inc..Walgreens sunscreen lotion (SPF 30,, Avobenzone 3.0%, Homosalate 10.0%, Octisalate 5.0%, Octocrylene 10%) was purchased from Walgreens. Sunscreen oil (SPF 30, Avobenzone 3.0%, Homosalate 10.0%, Octisalate 5.0%, Octocrylene 7%) was from L'Oréal Paris.
10
Chitosan-based Nanoparticle Drug Delivery
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All chemicals were purchased from respective companies (in brackets) and were used without pretreatment or purification. Pyridine, absolute ethanol, and acetone of analytical grades (Carlo Erba France, and Labchem, USA). Medium molecular weight chitosan, phenylsuccinic anhydride, and sodium hexametaphosphate (HMP) (Sigma-Aldrich, USA). Polyphosphoric Acid (PPA) and phthalic anhydride (Fluka, Switzerland). Ultrapure water (conductivity = 0.05 μs/cm) for DLS size analysis (Millipore, USA).
Penta basic sodium tripolyphosphate (Sigma-Aldrich, Germany), N-ethyl-N′-(3-dimethylaminopropyl) carbodiimide hydrochlorides (EDC) (Sigma- Aldrich, USA), hydrochloric acid (37%) (Carlo Erba, Spain) and sodium hydroxide (Rasayan Laboratory, India). Dialysis tubing (molecular weight cutoff = 14 kDa, Sigma-Aldrich, USA), Tris base buffer (Bio Basic Inc., Canada), Methylene blue (Seelze, Germany), and Doxorubicin HCl (Ebwe Pharma, Austria). CellTiter Non-Radioactive Cell Proliferation Assay Kit from Promega (USA). RPMI 1640 medium and fetal bovine serum (FBS) were purchased from (Caissan, USA), L-glutamine, penicillin–streptomycin and trypsin-EDTA were purchased from (EURO Clone, Italy). 4′,6-Diamidino-2-phenylindole (DAPI) stain was purchased from Invitrogen (Thermo Fisher Scientific, USA). Poly-L-lysine obtained from (Sigma- Aldrich, Germany).
Penta basic sodium tripolyphosphate (Sigma-Aldrich, Germany), N-ethyl-N′-(3-dimethylaminopropyl) carbodiimide hydrochlorides (EDC) (Sigma- Aldrich, USA), hydrochloric acid (37%) (Carlo Erba, Spain) and sodium hydroxide (Rasayan Laboratory, India). Dialysis tubing (molecular weight cutoff = 14 kDa, Sigma-Aldrich, USA), Tris base buffer (Bio Basic Inc., Canada), Methylene blue (Seelze, Germany), and Doxorubicin HCl (Ebwe Pharma, Austria). CellTiter Non-Radioactive Cell Proliferation Assay Kit from Promega (USA). RPMI 1640 medium and fetal bovine serum (FBS) were purchased from (Caissan, USA), L-glutamine, penicillin–streptomycin and trypsin-EDTA were purchased from (EURO Clone, Italy). 4′,6-Diamidino-2-phenylindole (DAPI) stain was purchased from Invitrogen (Thermo Fisher Scientific, USA). Poly-L-lysine obtained from (Sigma- Aldrich, Germany).
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