The following antibodies were employed: myosin heavy chain (sc-376157), myoD (sc-760), Pax-7 (sc-81975), β-actin HRP-conjugated antibody (sc-47778HRP), and GAPDH (sc-26778) from Santa Cruz Biotechnology (Dallas, TX, USA); collagen I (Ab21286) and collagen IV (ab19808) from Abcam, Cambridge, UK; utrophin (NCL-DRP2) from Leica Biosystems Inc. (Buffalo Grove, IL, USA); donkey anti-rabbit Cy-3-conjugated antibody (#711–166–152) and donkey anti-mouse Cy-3-conjugated antibody (#715–165–151) from Jackson ImmunoResearch Laboratories Inc. (West Grove, PA, USA); goat anti-mouse Alexa488-conjugated antibody (#A-11029) from Molecular Probes Inc., Carlsbad, CA, USA; and 4′,6-diamidino-2-phenylindole (DAPI) (D1306) from ThermoFischer Scientific (Oregon City, OR, USA).
4 6 diamidino 2 phenylindole dapi d1306
Manufactured by Thermo Fisher Scientific
Sourced in United States
4,6-diamidino-2-phenylindole (DAPI-D1306) is a fluorescent stain that binds to DNA. It is commonly used in various biological and laboratory applications.
Automatically generated - may contain errors
Lab products found in correlation
Alexa 488 conjugated goat anti mouse antibody, by Thermo Fisher Scientific (1 mentions)
Gapdh, by Santa Cruz Biotechnology (1 mentions)
Cryostat microtome, by Leica (1 mentions)
A1si laser scanning confocal microscope, by Nikon (1 mentions)
Nis elements ar analysis 4, by Nikon (1 mentions)
Vectashield mounting medium h 1000, by Vector Laboratories (1 mentions)
Streptozocin, by Merck Group (1 mentions)
Fibronectin 610077, by BD (1 mentions)
L glucose, by Merck Group (1 mentions)
D glucose, by Merck Group (1 mentions)
Ferric chloride hexahydrate, by Thermo Fisher Scientific (1 mentions)
Migration assay kit, by Merck Group (1 mentions)
Ammonium hydroxide, by Thermo Fisher Scientific (1 mentions)
A549 ccl 185, by American Type Culture Collection (1 mentions)
Apoptosis and necrosis quantification kit, by Biotium (1 mentions)
Hydrochloric acid (hcl), by Thermo Fisher Scientific (1 mentions)
Mes sodium salt, by Thermo Fisher Scientific (1 mentions)
N n dimethylformamide (dmf), by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
1 ethyl 3 3 dimethylaminopropyl carbodiimide hydrochloride edc, by Thermo Fisher Scientific (1 mentions)
13 protocols using 4 6 diamidino 2 phenylindole dapi d1306
1
Immunofluorescence Analysis of Skeletal Muscle
2
Zebrafish Pancreatic β-cell Ablation Imaging
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Fluorescence imaging of the transgenic zebrafish was conducted to identify the ablation of β-cells in the pancreas. The pancreases of zebrafish were fixed overnight in 4% paraformaldehyde. For histological analysis, the fixed samples were embedded in 1.5% agar blocks containing 5% sucrose. Frozen blocks were sectioned into 10 μm thick slices using a cryostat microtome (Leica, Wetzlar, Germany), and the sectioned slices were rinsed with phosphate-buffered saline (PBS). Sectioned slices were stained with 4′,6-diamidino-2-phenylindole (DAPI, D1306, Thermo Fisher Scientific, Waltham, MA, USA) to counterstain the cells. All fluorescence images were captured using an A1Si laser-scanning confocal microscope (Nikon, Tokyo, Japan). Confocal images (1 μm z-stacking) were processed using NIS-Elements AR Analysis 4.30 software (Nikon).
3
Immunostaining of Gastrula-stage Embryos
4
Immunoblotting and Immunofluorescence Markers
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L-glucose, D-glucose, and streptozocin were purchased from Sigma (St. Louis, MO, USA). α-SMA (A2547), β-actin (A5411), and Twist (T6451) antibodies were purchased from Sigma-Aldrich (St Louis, MO, USA). Antibodies against Vimentin (550513), N-cadherin (610920), and fibronectin (610077) were obtained from BD (Franklin Lakes, NJ, USA). Antibodies for Snail (3879S), β-catenin (9582S), and Glial Fibrillary Acidic Protein (GFAP, 3670S) were from Cell Signaling Technology (Danvers, MA, USA). CTGF (ab6922) and Glutamine synthase (GS, ab176562) antibodies were purchased from Abcam (Cambridge, MA, USA). Glutamine synthase (GS, MAB302) antibody was from Millipore (Billerica, MA, USA). Goat Anti-mouse(PI-2000) and anti-rabbit (PI-1000) horseradish peroxidase (HRP)-conjugated secondary antibodies were from Vector Laboratories (Burlingame, CA, USA). Alexa Flour 488 goat anti-rabbit/anti-mouse (A21206/A21202) and Alexa Fluor 594 goat anti-rabbit/anti-mouse (A21207/A21203) antibodies and 4’,6-Diamidino-2-phenylindole (DAPI, D1306) were from Life Technology (St. Louis, MO, USA).
5
Synthesis and Characterization of Nanoparticles
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Ferric chloride hexahydrate (FeCl3.6H2O), ferrous chloride tetrahydrate (FeCl2.4H2O), hydrochloric acid and ammonium hydroxide were obtained from Fischer Scientific. DMF, DMSO, 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), N-hydroxysuccinimide (NHS), polyacrylic acid (PAA), propargylamine (PA) and other chemicals were purchased from Sigma-Aldrich. Optical dye (DiI-D282), dihydroethidium (DHE), mitosox red and 4,6-diamidino-2-phenylindole (DAPI-D1306) were purchased from Invitrogen, whereas the (1-ethyl-3-[3-(dimethylamino) propyl] carbodiimide hydrochloride) (EDC) was obtained from Pierce Biotechnology. MES sodium salt was purchased from Acros organic. Apoptosis and necrosis quantification kit was purchased from Biotium and the migration assay kit was purchased from Millipore. The human lung carcinoma cell line A549 (CCL-185) and Chinese hamster cells (CHO) were obtained from ATCC. Magnetic column QuadroMACSTM LS was obtained from Miltinyi Biotec and the dialysis membrane was purchased from spectrum laboratories. Dulbecco's modified eagle (DMEM) medium and Kaighn's modification of Ham's F12K medium from Corning.
6
Comprehensive Immunohistochemical Analysis
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Antibodies used in the study are: αSMA (ab7817) and Ki67 (ab15580) from Abcam; GM130 (610822) from BD Biosciences; cleaved caspase-3 (9664S) and PDGFRα (3174S) from Cell Signaling Technology; BrdU (GTX26326) from GeneTex; pro-surfactant protein C (10774–1-AP) from Proteintech; caveolin-1 (sc-894), Hopx (sc-398,703) and GAPDH (sc25778) from Santa Cruz Biotechnology; β-actin (A5441) and α-tubulin (T5168) from Sigma-Aldrich; and AlexaFluor 488, 594 or 647-conjugated secondary antibodies from Invitrogen. Chemicals used in the study are: ProLong antifade (P36930) and 4′,6-diamidino-2-phenylindole (DAPI, D1306) reagents from Invitrogen; and Avertin (T48402), methacholine (A2251) and 5-bromo-2′-deoxyuridine (B9285) from Sigma-Aldrich.
7
Colonic Histological and Immunofluorescence Analysis
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The collected colon was washed three times with sterile PBS. A part of the colon tissue was then fixed with 10% phosphate-buffer formalin for 24 h. After fixation, the tissue sample was dehydrated through an ethanol series, followed by embedding in paraffin. The paraffin blocks were then sectioned (5 µm) and stained with hematoxylin-eosin for histological evaluation. For mucosal layer evaluation, Alcian blue was used to stain the mucin in the paraffin-embedded sections and the nuclei were counterstained with nuclear fast red. For immunofluorescence analysis, the cut sections were stained with ZO-1 (#61-7300; Invitrogen, Carlsbad, CA, USA) or claudin-1 antibodies (#71-7800; Invitrogen) and goat anti-rabbit Alexa Flour 488 secondary antibody (A-11008; Invitrogen). 4′,6-diamidino-2-phenylindole (DAPI; D-1306; Invitrogen) was used to counterstain cell nuclei. Slides were examined and analyzed using an epifluorescence microscope.
8
Comprehensive Antibody Sourcing for Biochemical Experiments
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Phenylephrine (PE; 046K1351), angiotensin II (Ang II; A9525), and anti‐FLAG M2 monoclonal antibody (A8592, F1804) were purchased from Sigma–Aldrich (St. Louis, MO). Antibodies against Actin (sc‐1615) and donkey anti‐goat IgG (sc‐2020) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA) for Western blotting. Antibodies against phospho‐ERK1/2 (#9101S), phospho‐MEK (#9121S), MEK (#9122S), p‐c‐Raf (#9427S), c‐Raf (#9422), Histone H3 (#9715), α‐tubulin (#2144), and rabbit IgG (#7074S) were purchased from Cell Signaling Technology (Danvers, MA). The antibody against ERK1/2 (V114A) was purchased from Promega (Madison, WI). The antibody against Tom20 (612278) was purchased from BD Biosciences (San Jose, CA). The antibody against glyceraldehyde3‐phosphate dehydrogenase (Gapdh) (HAB374) was purchased from EMD Millipore (Billerica, MA). The antibody against α1 Na, K‐ATPase (ab7671) was purchased from Abcam (Cambridge, UK). An anti‐Mtus1 polyclonal antibody raised in rabbits targeted a peptide (CSPKRSPTSSAIPFQSPRNSGSFSSPSISPR) within the C terminus of the protein. The antibodies for immunofluorescence staining of Alexa mouse Fluor 488 (A11029), rabbit Fluor 488 (A11034), rabbit Fluor 546 (A11035), Fluor 568 phalloidin (A12380), and 4′,6‐diamidino‐2‐phenylindole (DAPI) (D1306) were purchased from Invitrogen (Carlsbad, CA).
9
Confocal Fluorescence of Viral Protein Localization
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For confocal fluorescence, the cells were grown on coverslips and then infected with PRV as indicated at an MOI of 10. At 2 and 10 hpi, the cells were fixed with 4% paraformaldehyde for 30 min. The fixed cells were permeabilized with 0.1% Triton X-100 in PBS for 15 min and blocked with 3% bovine serum albumin in PBS for 2 h. Anti-LC3 antibodies were added to detect LC3 expression for 2 h, then incubated with FITC-conjugated goat anti-rabbit secondary antibodies. The second anti-gE antibodies were added to the fixed cells to detect gE expression for 2 h, and TRITC-conjugated goat anti-mouse secondary antibodies were added next. The nuclei were stained with 4′-6-Diamidino-2-Phenylindole (DAPI, D1306, Invitrogen, U.S.A.). To detect co-localization of LC3 and SQSTM1, anti-LC3 antibodies and anti-SQSTM1 antibodies and their respective secondary antibodies were used as indicated.
10
Confocal Microscopy Immunofluorescence Assay
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Cells of interest grown on chamber slides were fixed and permeabilized. Following blocking with 10% goat and donkey serum (G9023 and D9663, Sigma-Aldrich), they were incubated with primary antibodies overnight at 4°C. The slides were washed and incubated with Alexa-Fluor®-488 donkey anti-mouse IgG and Alexa-Fluor®-568 goat anti-rabbit IgG (A21202 and A11011, Molecular Probes®, Invitrogen) together with 4′,6-Diamidino-2-Phenylindole (DAPI) (D-1306, Molecular Probes®) for nucleus staining. All images were captured using Zeiss LSM700 inverted confocal microscope (Gottingen, Germany) at Faculty Core Facility, Li Ka Shing Faculty of Medicine, the University of Hong Kong.
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