4 6 diamidino 2 phenylindole dihydrochloride

Manufactured by Merck Group

Sourced in United States, China, United Kingdom

4',6-diamidino-2-phenylindole dihydrochloride is a fluorescent dye that binds to DNA. It is commonly used in various biological and biochemical applications to stain and visualize nucleic acids.

Automatically generated - may contain errors

Lab products found in correlation

4 6 diamidino 2 phenylindole (dapi), by Merck Group (10 mentions) Bovine serum albumin (bsa), by Merck Group (4 mentions) Triton x 100, by Merck Group (3 mentions) Fluorescence microscope, by Leica camera (3 mentions) Tcs sp5, by Leica camera (3 mentions) Dimethyl sulfoxide (dmso), by Merck Group (3 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (3 mentions) Lsm 710 confocal microscope, by Zeiss (3 mentions) Confocal laser scanning microscope, by Leica Microsystems (2 mentions) Paraformaldehyde (pfa), by Merck Group (2 mentions) Cy3 conjugated goat anti mouse antibody, by Jackson ImmunoResearch (2 mentions) Prolong gold antifade reagent, by Thermo Fisher Scientific (2 mentions) Fv1000, by Olympus (2 mentions) Mitotracker red cmxros, by Thermo Fisher Scientific (2 mentions) 3 4 5 dimethylthiazol 2 yl 2 5 diphenyltetrazolium bromide mtt, by Merck Group (2 mentions) Crl 1658, by American Type Culture Collection (2 mentions) Fluorescence microscope, by Nikon (2 mentions) Formaldehyde solution, by Merck Group (2 mentions) Lysotracker deep red, by Thermo Fisher Scientific (2 mentions) Phosphate buffered saline (pbs), by Thermo Fisher Scientific (2 mentions)

86 protocols using 4 6 diamidino 2 phenylindole dihydrochloride

Visualizing MyoG Expression in Transfected MSCs

Check if the same lab product or an alternative is used in the 5 most similar protocols

The MyoG-EGFP construct encoded both MyoG and enhanced green fluorescent protein. The
coding sequence of the rat MyoG gene (GenBank Accession No. NM_017115) was subcloned
into the Xho I and Kpn I sites of the plasmid
vector GV143 (Shanghai Genechem). At 80% confluence, cells were transfected using
X-tremeGENE HP DNA transfection reagent (Roche, Switzerland). Cells transfected with
the empty vector were used as negative controls to determine the MyoG-specific
transfection efficacy. Laser scanning confocal microscopy (BX61W1-FV1000; Olympus,
Japan) was used to detect immunofluorescence in MyoG-transfected MSCs (MTMs). A mouse
anti-MyoG monoclonal antibody (M5815; Sigma-Aldrich, USA) was used at a 1:100
dilution for immunostaining. Nuclear staining was performed using
4',6'-diamidino-2-phenylindole dihydrochloride (100 nM; Sigma).

Shen H., Lv Y., Shen X.Q., Xu J.H., Lu H., Fu L.C, & Duan T. (2016). Implantation of muscle satellite cells overexpressing myogenin improves denervated muscle atrophy in rats. Brazilian Journal of Medical and Biological Research, 49(2), e5124.

+ Open protocol

+ Expand

Optimizing Transfection Efficiency for Gene Delivery

Check if the same lab product or an alternative is used in the 5 most similar protocols

Materials used in this study were l-histidine hydrochloride, l-arginine, stearic acid, and hydrogen peroxide (30% w/v; Sangon Biotech, Shanghai, People’s Republic of China); polyethylenimine (PEI, branched, molecular weight 25 kDa), Lipofectamine™ 2000, l-buthionine-sulfoximine, 4′,6-diamidino-2-phenylindole dihydrochloride, l-cysteine hydrochloride monohydrate, ethidium bromide, dithiothreitol (DTT, Sigma-Aldrich, St Louis, MO, USA); a luciferase assay kit (Promega, Madison, WI, USA); pDNA (pGL3 and pEGFP) (Shanghai Innovation Biotechnology Co Ltd, Shanghai, People’s Republic of China); enhanced bicinchoninic acid protein assay kit (Beyotime, Nanjing, People’s Republic of China); Dulbecco’s Modified Eagle’s Medium (DMEM), fetal bovine serum, YOYO-1 (Y3601) and penicillin–streptomycin solution 5 kU/mL (Life Technologies, Carlsbad, CA, USA); a Cell Counting Kit-8 (CCK-8, Dojindo Molecular Technologies Inc, Nanjing, People’s Republic of China); and HEK293 cells and HeLa cells (Cell Culture Center of the Shanghai Institutes for Biological Sciences of the Chinese Academy of Sciences, Shanghai, People’s Republic of China). All other reagents were of analytical grade. All animal experiments were performed in accordance with the ethics and regulations of animal experiments of Second Military Medical University (Shanghai, People’s Republic of China).

Yao C., Tai Z., Wang X., Liu J., Zhu Q., Wu X., Zhang L., Zhang W., Tian J., Gao Y, & Gao S. (2015). Reduction-responsive cross-linked stearyl peptide for effective delivery of plasmid DNA. International Journal of Nanomedicine, 10, 3403-3416.

+ Open protocol

+ Expand

In Vivo Tracking of PKH26-Labeled hMSCs

Check if the same lab product or an alternative is used in the 5 most similar protocols

Within 4 weeks of PKH26-hMSC transplantation, frozen sections were extracted from kidney tissues weekly. Successive frozen sections (thickness, 5 µm) were made following Optimal Cutting Temperature (OCT) compound embedding. Subsequent to drying for 30 min at room temperature, the sections were fixed with cold acetone for 10 min. An Olympus IX70 fluorescence microscope (Olympus Corporation, Tokyo, Japan) and a laser scanning confocal microscope (Leica Microsystems, Inc., Buffalo Grove, IL, USA) were used to observe and analyze the sections.
The localization of PKH26-labeled hMSCs in the kidneys was observed 2 weeks after the injection of cells. Samples were frozen immediately in liquid nitrogen, embedded in OCT compound, sliced to 5 µm sections, fixed in acetone for 10 min, and incubated for 30 min at 22°C with FITC-labeled wheat germ agglutinin (WGA; Vector Laboratories, Inc.). Nuclei were stained with 4,6-diamidino-2-phenylindole dihydrochloride (Sigma-Aldrich). PKH26-positive cells were counted in 10 frozen renal sections per mouse (n=3 mice). The number of stem cells in kidney tubules was also counted.

Yuan L., Liu H.Q, & Wu M.J. (2016). Human embryonic mesenchymal stem cells participate in differentiation of renal tubular cells in newborn mice. Experimental and Therapeutic Medicine, 12(2), 641-648.

+ Open protocol

+ Expand

Polymer Synthesis and Transfection Reagents

Check if the same lab product or an alternative is used in the 5 most similar protocols

Lipofectamine 2000 (Invitrogen, Carlsbad, CA), Opti-MEM I (Invitrogen), pEGFP-N1 DNA (Elim Biopharmaceuticals, Hayward, CA), pDsRed-Max-N1^{39 (link)} (Addgene DNA plasmid 21718, Cambridge, MA), and cell culture media components were used as received. Monomers used for synthesizing polymers (Scheme 1) were purchased as follows: 1,3-butanediol diacrylate (B3b; Sigma-Aldrich, St. Louis, MO); 1,4-butanediol diacrylate (B4; Alfa Aesar, Ward Hill, MA); 1,5-pentanediol diacrylate (B5; Monomer-Polymer and Dajac Laboratories, Trevose, PA); 3-amino-1-propanol (S3; Alfa Aesar); 4-amino-1-butanol (S4; Alfa Aesar); 5-amino-1-pentanol (S5; Alfa Aesar); 1,3-diaminopropane (E1; Sigma-Aldrich); 1,3-diaminopentane (E3; TCI America, Portland, OR); 2-methyl-1,5-diaminopentane (E4; TCI America); 1,11-diamino-3,6,9-trioxaundecane (E5; TCI America); 2-(3-aminopropylamino)ethanol (E6; Sigma-Aldrich); 1-(3-aminopropyl)-4-methylpiperazine (E7; Alfa Aesar); 1-(3-aminopropyl)pyrrolidine (E8; TCI America); and cystamine dihydrochloride (E10; Alfa Aesar). 4′,6-Diamidino-2-phenylindole dihydrochloride (DAPI) was purchased from Sigma and used as a 750 nM solution in PBS. Other materials were reagent grade.

Guerrero-Cázares H., Tzeng S.Y., Young N.P., Abutaleb A.O., Quiñones-Hinojosa A, & Green J.J. (2014). Biodegradable Polymeric Nanoparticles Show High Efficacy and Specificity at DNA Delivery to Human Glioblastoma in Vitro and in Vivo. ACS Nano, 8(5), 5141-5153.

+ Open protocol

+ Expand

Immunolabeling of EBOV-infected ARPE-19 Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Confluent monolayers of fixed EBOV-infected ARPE-19 cells were permeabilized with 0.1% Nonidet P-40 (Sigma-Aldrich, St. Louis, MO) for 10 minutes and blocked with PBS 1% bovine serum albumin (Sigma-Aldrich) for 90 minutes. Cells were incubated overnight at room temperature with rabbit anti-Ebolavirus nucleoprotein antiserum,^{19 (link)} diluted 1:200 in blocking solution. Subsequently, cells were washed three times with PBS with 0.05% Tween 20 (PBS-T), and incubated with Alexa Fluor 488-conjugated goat anti-rabbit secondary antibody (Thermo Fisher Scientific-Molecular Probes, Eugene, OR) at 2 μg/mL in blocking solution for 60 minutes. Finally, monolayers were washed three times with PBS-T, treated with 0.1 μg/mL nM 4′6-diamidino-2-phenylindole-dihydrochloride (Sigma-Aldrich) in PBS for 10 minutes, and washed three times in PBS-T and two times in PBS. Immunolabeled ARPE-19 cells were imaged on the EVOS FL Cell Imaging System (Thermo Fisher Scientific-Invitrogen) at ×10 magnification. Mock-infected monolayers of ARPE-19 cells were immunolabeled and imaged in parallel as control.

Smith J.R., Todd S., Ashander L.M., Charitou T., Ma Y., Yeh S., Crozier I., Michael M.Z., Appukuttan B., Williams K.A., Lynn D.J, & Marsh G.A. (2017). Retinal Pigment Epithelial Cells are a Potential Reservoir for Ebola Virus in the Human Eye. Translational Vision Science & Technology, 6(4), 12.

+ Open protocol

+ Expand

Multi-color FISH Assay for Pituitary Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Multi-color FISH was performed as previously described in (35 (link)) with minor modifications. Tissues were serially rehydrated, and the pituitary was detached from the brain. Afterwards, whole pituitaries were hybridized with the probes (0.11 – 3.17 ng/µl) for 18 hours at 55°C, and incubated with different combinations of anti-DNP- (Perkin Elmer), anti-FITC-, and anti-DIG-conjugated antibodies (Roche Diagnostics), followed by TAMRA- (Thermofisher), Cy5- (Perkin Elmer) and FITC-conjugated tyramides (Sigma). The nuclei were stained with DAPI (1:1000, 4’, 6-diamidino-2-phenylindole dihydrochloride; Sigma). The absence of labeling when using sense probes was used to confirm the specificity of the anti-sense probes. Whole pituitaries were mounted using Vectashield H-1000 Mounting Medium (Vector, Eurobio/Abcys) between microscope slides and cover slips (Menzel Glässer, VWR) with spacers (Reinforcement rings, Herma) in between for the juveniles, and between two cover slips with spacers for adults.

Royan M.R., Siddique K., Csucs G., Puchades M.A., Nourizadeh-Lillabadi R., Bjaalie J.G., Henkel C.V., Weltzien F.A, & Fontaine R. (2021). 3D Atlas of the Pituitary Gland of the Model Fish Medaka (Oryzias latipes). Frontiers in Endocrinology, 12, 719843.

+ Open protocol

+ Expand

Cytotoxicity Evaluation of Aspirin and 5-FU

Check if the same lab product or an alternative is used in the 5 most similar protocols

The analyzed compounds (aspirin and 5-fluorouracil) and the other reagents used in the present study dimethyl sulfoxide (DMSO), fetal calf serum (FCS), penicillin/streptomycin, trypsin-EDTA solution, phosphate saline buffer (PBS), MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide), 4′,6-Diamidino-2-phenylindole dihydrochloride, and 2-(4-Amidinophenyl)-6-indolecarbamidine dihydrochloride (DAPI), were purchased from Sigma Aldrich, Merck KgaA (Darmstadt, Germany), and Alexa Fluor^® 555 Phalloidin was acquired from Cell Signaling USA.
Cell lines were cultured in the specific media DMEM (P04-03550) and McCoy’s 5A (P04-05500), which were purchased from PAN Biotech GmbH (Aidenbach, Germany). As part of RT-PCR, the following primers were used: 18S, Bax, Bcl-2 purchased from Thermo Fisher Scientific, Inc. (Waltham, MA, USA) and Bad, caspase 3, and caspase 8, purchased from Eurogentec (Seraing, Belgium). All reagents presented appropriate characteristics for use in cell culture.

Susan M., Macasoi I., Pinzaru I., Dehelean C., Ilia I., Susan R, & Ionita I. (2023). In Vitro Assessment of the Synergistic Effect of Aspirin and 5-Fluorouracil in Colorectal Adenocarcinoma Cells. Current Oncology, 30(7), 6197-6219.

+ Open protocol

+ Expand

Immunohistochemical Analysis of Cardiac Tissue

Check if the same lab product or an alternative is used in the 5 most similar protocols

Immunohistochemical staining was conducted on 4‐mm sections of heart tissue using Mff 1:500 (Abcam), phospho‐endothelial nitric oxide synthase (p‐eNOS; Ser1177) 1:200 (Abcam), intercellular adhesion molecule–1 1:500 (Abcam), vascular cell adhesion molecule–1 1:500 (Abcam), F4/80 1:500 (Abcam), and plasma albumin 1:500 (Abcam). The primary antibodies were as follows: CD31 (1:1500, Abcam), vascular endothelial cadherin (VE‐cadherin; 1:1000, Abcam), cyt‐c (1:500, Abcam), Mff (1:1000, Abcam), pDrp1 (1:500, Abcam), HK2 (1:500, Abcam), cleaved caspase3 (CL.caspase3; 1:1000, Abcam), and pro‐caspase3 (1:2000, Abcam). 4′,6‐Diamidino‐2‐phenylindole dihydrochloride (Sigma‐Aldrich, USA), lysosome stain, and a mitochondrion‐selective MitoFluor stain (Molecular Probes, USA) were used to marker the nuclear, lysosome, and mitochondria, respectively. For the cross‐linking of VDAC1, treated cells were harvested and added with dimethyl sulfoxide as a vehicle control (2%, as used in compound‐containing samples) or cross‐linked with 0.5 mm ethylene glycolbis (succinimidyl succinate) for 10 minutes at 30°C followed by 20 mm Tris‐HCl (pH 7.4) to quench the reaction. Samples were determined by SDS‐PAGE via immunoblotting.

Zhou H., Hu S., Jin Q., Shi C., Zhang Y., Zhu P., Ma Q., Tian F, & Chen Y. (2017). Mff‐Dependent Mitochondrial Fission Contributes to the Pathogenesis of Cardiac Microvasculature Ischemia/Reperfusion Injury via Induction of mROS‐Mediated Cardiolipin Oxidation and HK2/VDAC1 Disassociation‐Involved mPTP Opening. Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease, 6(3), e005328.

+ Open protocol

+ Expand

Hyaluronic Acid-Based Theranostic Nanoparticles

Check if the same lab product or an alternative is used in the 5 most similar protocols

Hyaluronic acid (HA, Mw = 4.8 kDa), 3-(diethylamino)propylamine (DEAP), N-hydroxysuccinimide (NHS), N,N’-dicyclohexylcarbodiimide (DCC), triethylamine (TEA), deoxycholic acid (DOCA), 4-dimethylamino pyridine (DMAP), pyridine, dimethyl sulfoxide (DMSO), sodium tetraborate, adipic acid dihydrazide (ADH), doxorubicin hydrochloride (DOX), paraformaldehyde, heparin, and Triton X-100, 4′,6-diamidino-2-phenylindole dihydrochloride (DAPI) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Chlorin e6 (Ce6) was purchased from Frontier Scientific Inc (Logan, UT, USA). RPMI-1640 medium, DMEM medium, fetal bovine serum (FBS), phosphate buffered saline (PBS), ethylene diamine tetra-acetic acid (EDTA), penicillin, trypsin, and streptomycin were purchased from Welgene Inc (Seoul, Korea). EV-depleted FBS was purchased from System Biosciences Inc. (Palo Alto, CA, USA). Cell Counting Kit-8 (CCK-8) was purchased from Dojindo Molecular Technologies Inc. (Santa Clara, CA, USA). Wheat Germ Agglutinin Alexa Fluor^® 488 Conjugate (WGA-Alexa Fluor^® 488), fluorescein isothiocyanate (FITC) were purchased from Life Technologies (Carlsbad, CA, USA).

Park J., Lee H., Youn Y.S., Oh K.T, & Lee E.S. (2020). Tumor-Homing pH-Sensitive Extracellular Vesicles for Targeting Heterogeneous Tumors. Pharmaceutics, 12(4), 372.

+ Open protocol

+ Expand

Confocal Microscopy Evaluation of HMGB1 Translocation in SAH

Check if the same lab product or an alternative is used in the 5 most similar protocols

Confocal microscopy was used to observe HMGB1 translocation after SAH. Brain sections were prepared as described in the immunohistochemistry. Following rehydration, sections were incubated with 0.2% Triton X-100 in PBS for 5 min at room temperature and washed with PBS 3 times for 5 min. Sections were placed in 10 mmol/l citrate buffer (pH 6.0) and heated in the microwave oven at 95°C for 30 min. Sections were cooled at room temperature for 20 min and rinsed in PBS. Nonspecific protein binding was blocked by incubation in 5% bovine serum for 40 min. Sections were incubated with primary rabbit anti-HMGB1 (1:200; ab190377; Abcam) overnight at 4°C, followed by washing with PBS 3 times for 5 min. Sections were incubated with goat anti-rabbit secondary antibody (1:100) conjugated with fluorescein isothiocyanate and tetramethylrhodamine (cat. no. ZF-0311; Beijing Zhongshan Golden Bridge Biotechnology, Beijing, China) at 37°C for 1 h. The cell nucleus was stained with 4,6′-diamidino-2-phenylindole, dihydrochloride (1:1,000; Sigma-Aldrich; Merck KGaA). Immunofluorescence was detected using a laser scanning confocal microscope (Olympus FV10-ASW; Olympus Corp., Tokyo, Japan).

An J.Y., Pang H.G., Huang T.Q., Song J.N., Li D.D., Zhao Y.L, & Ma X.D. (2017). AG490 ameliorates early brain injury via inhibition of JAK2/STAT3-mediated regulation of HMGB1 in subarachnoid hemorrhage. Experimental and Therapeutic Medicine, 15(2), 1330-1338.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!