CTAC, TEA, xanthine, and xanthine oxidase (XO) were purchased from Sigma-Aldrich. TEOS and NaOH were obtained from Shanghai Lingfeng Chemical Reagent Co., Ltd. Fe(acac)3 was provided by J&K Scientific. Urea, RhB, iron chloride hexahydrate (FeCl3·6H2O) and 3,3′,5,5′-tetramethyl-benzidine (TMB) were bought from Sinopharm Chemical Reagents Co., Ltd. Methoxy PEG silane was purchased from Shanghai Yare Biotech, Co., Ltd. Gallic acid was acquired from Macklin. Dulbecco’s modified eagle’s medium (DMEM), fetal bovine serum (FBS), penicillin/streptomycin, and PBS were provided by Gibco. FITC, 4′, 6-diamidino-2-phenylindole (DAPI), DCFH-DA, CCK-8 assay, calcein-AM, PI, annexin V-FITC, H&E, paraformaldehyde (PFA) and TUNEL apoptosis assay kit were acquired from Beyotime. Primary antibody against GPX4 and a biotinylated secondary antibody (anti-rabbit IgG, HRP-linked antibody) were purchased from Cell Signaling Technology.
Paraformaldehyde pfa
Manufactured by Beyotime
Sourced in China
Paraformaldehyde (PFA) is a white, crystalline solid that is commonly used as a fixative in various laboratory applications. It is the polymerized form of formaldehyde and serves as a source of formaldehyde. PFA is known for its ability to preserve the structural integrity and morphology of biological samples during preparation for microscopy and other analytical techniques.
Automatically generated - may contain errors
Lab products found in correlation
Gallic acid, by Maclin Biochemical Technology (1 mentions)
Xanthine, by Merck Group (1 mentions)
Xanthine oxidase, by Merck Group (1 mentions)
Fe acac 3, by J&K Scientific (1 mentions)
Iron chloride hexahydrate fecl3 6h2o, by Sinopharm (1 mentions)
3 3 5 5 tetramethyl benzidine tmb, by Sinopharm (1 mentions)
Anti rabbit igg hrp linked antibody, by Cell Signaling Technology (1 mentions)
Tunel apoptosis assay kit, by Beyotime (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Phosphate buffered saline (pbs), by Thermo Fisher Scientific (1 mentions)
Calcein am, by Beyotime (1 mentions)
Annexin 5 fitc, by Beyotime (1 mentions)
Cck 8 assay, by Beyotime (1 mentions)
Urea, by Sinopharm (1 mentions)
Sodium hydroxide (naoh), by Shanghai Lingfeng Chemical Reagent (1 mentions)
Dapi solution, by Solarbio (1 mentions)
Tunel staining solution, by Roche (1 mentions)
Phosphate buffered saline (pbs), by Solarbio (1 mentions)
Phosphate buffered saline (pbs), by Beyotime (1 mentions)
Hematoxylin, by Merck Group (1 mentions)
5 protocols using paraformaldehyde pfa
1
Multifunctional Nanomaterial Synthesis and Evaluation
2
Quantifying Apoptosis in BMSCs
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To measure the apoptosis levels of BMSCs, about 1 × 105 cells were plated in a small dish. The cells treated with Res were used for terminal deoxynucleotidyl transferase dUTP nick‐end labeling (TUNEL) staining. The culture medium in the dish was discarded, and the cells were fixed in 4% Paraformaldehyde (PFA) (Beyotime) after washed with PBS (Beyotime). After the closure and penetration procedures, TUNEL staining solution (Roche, Basel City, Switzerland) Vial1 and Vial2 were mixed in a ratio of 1:9 according to the instructions of the kit. Then, cells were incubated by the mixture for 1 hour in a dark room. After washed with PBS, cells were incubated with DAPI solution (Solarbio) for 30 minutes. Finally, the cells were observed under a fluorescence microscope (Nikon Corporation, Tokyo City, Japan).
3
Immunohistochemical and Immunofluorescence Analysis of KI-67 and Immune Markers
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For immunohistochemical (IHC) analysis of KI-67, tumor tissues were fixed with 4% paraformaldehyde (PFA; Beyotime, Shanghai, China), followed by treatment with 0.1% hydrogen peroxide (Sigma, St Louis, USA), embedding in paraffin and sectioning. The slides were blocked with 10% goat serum in PBS solution and incubated with anti-KI-67 antibody (27309-1-AP; Proteintech, Wuhan, China) overnight at 4°C in a wet box. After incubation with the secondary antibody for 2 h, the samples were treated with an ABC HRP kit (Yeasen, Shanghai, China) according to the manufacturer’s instructions. Then, DAB (3,3-diaminobenzidine) HRP substrate was used before counterstaining with hematoxylin (Sigma) and dehydration.
For immunofluorescence (IF) microscopic analysis, after being blocked with 10% goat serum in PBS solution, the slides were incubated with anti-CD8 antibody (66868-1-Ig; Proteintech) or anti-IFN antibody (15365-1-AP; Proteintech), followed by incubation with AF647 fluorescently labelled secondary antibodies (4410, 4414; Cell Signaling Technology, Danvers, USA) in the dark. Samples were stained with DAPI for 15 min and mounted before image acquisition. Photomicrographs were taken with a Zeiss imaging platform (LSM980; Zeiss, Jena, Germany) and SPOT Basic software. The IHC and IF images were analyzed by Image-Pro Plus software.
For immunofluorescence (IF) microscopic analysis, after being blocked with 10% goat serum in PBS solution, the slides were incubated with anti-CD8 antibody (66868-1-Ig; Proteintech) or anti-IFN antibody (15365-1-AP; Proteintech), followed by incubation with AF647 fluorescently labelled secondary antibodies (4410, 4414; Cell Signaling Technology, Danvers, USA) in the dark. Samples were stained with DAPI for 15 min and mounted before image acquisition. Photomicrographs were taken with a Zeiss imaging platform (LSM980; Zeiss, Jena, Germany) and SPOT Basic software. The IHC and IF images were analyzed by Image-Pro Plus software.
4
Electron Microscopy Sample Preparation
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The four above-mentioned groups of cells were cultured in 60 mm plates, collected in a phosphate-buffered saline (PBS) solution, and fixed with 2% (v/v) paraformaldehyde (PFA, Beyotime, Shanghai, China) containing 2.5% (w/v) glutaraldehyde (Sigma, St. Louis, MO, USA) buffered in Hank’s modified salt solution (HMSS, Procell, Wuhan, China) at 4 °C for 4 h. The cells were further fixed in a 1% (w/v) OsO4 solution buffered by 0.1 M cacodylate (pH 7.2) at 4 °C for 2 h. Then, the cells were scraped off the plastic and dehydrated with dinethanol. Dehydration was conducted in propylene oxide. The specimens were embedded in an EPON medium and cut into 60–70 nm sections. The samples were analyzed and recorded with a JEOL 1200 electron microscope (JEOL Ltd., Tokyo, Japan).
5
Cell Viability Assay and Colony Formation
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Cell viability was measured with a CCK-8 (Dojindo Laboratories, Kumamoto, Japan). HT1080 and Hepg2 cells were cultured overnight in a 96-well plate (Yongjin, Guangzhou, China) at densities of 2 × 103/well and 4 × 103/well. The cells were subjected to the preceding agents for 48 h and then 90 µL fresh medium and 10 µL CCK-8 solution (EdU; 10 µM) were added to them. Optical densities (OD) were measured in a microplate reader (BioTek Instruments, Winooski, VT, USA) at 450 nm. The EdU images were obtained by fluorescence microscopy, magnification 10x (Leica, Germany). Colony formation assays were used to evaluate cell viability. About 1,000 cells were placed in six-well plates and cultured in complete DMEM. The latter was changed every 2 d. After 1 wk, the colonies were washed twice with phosphate-buffered saline (PBS), incubated with 4% (v/v) paraformaldehyde (PFA; Beyotime Biotechnology, Shanghai, China) for 30 min, and stained with crystal violet solution (Beyotime Biotechnology).
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