Antibodies for p65, p-p65, BDNF, claudin-5, occludin, caludin-1, iNOS, COX-2, and β-actin were purchased from Cell Signaling Technology (Beverly, MA). Enzyme-linked immunosorbent assay (ELISA) kits for cytokines were purchased from Ebioscience (San Diego, CA). 4′,6-Diamidino-2-phenylindole dilactate (DAPI) was purchased from Sigma (St. Louis, MO). QIAamp Fast DNA stool mini kit was purchased from Qiagen (Hilden, Germany). Limulus amoebocyte lysate (LAL) assays was purchased from Cape Cod Inc. (East Falmouth, MA). General anaerobic medium (GAM), BL, and DHL media were from Nissui Pharmaceutical Co. (Tokyo, Japan). MRS medium was purchased from BD (Radnor, PA).
4 6 diamidino 2 phenylindole dilactate dapi
Manufactured by Merck Group
Sourced in United States, United Kingdom
4',6-diamidino-2-phenylindole dilactate (DAPI) is a fluorescent stain that binds strongly to DNA. It can be used to visualize and quantify DNA in a variety of biological applications.
Automatically generated - may contain errors
Lab products found in correlation
Alexa fluor 488, by Thermo Fisher Scientific (6 mentions)
Qiaamp fast dna stool mini kit, by Qiagen (5 mentions)
Alexa fluor 594, by Thermo Fisher Scientific (4 mentions)
Lipopolysaccharides (lps), by Merck Group (4 mentions)
Antibody for nf κb, by Cell Signaling Technology (4 mentions)
Lsm 710, by Zeiss (4 mentions)
Claudin 5, by Cell Signaling Technology (2 mentions)
Occludin, by Cell Signaling Technology (2 mentions)
Caludin 1, by Cell Signaling Technology (2 mentions)
Enzyme linked immunosorbent assay elisa kits for cytokines, by Thermo Fisher Scientific (2 mentions)
Cox 2, by Cell Signaling Technology (2 mentions)
β actin, by Cell Signaling Technology (2 mentions)
Mrs medium, by BD (2 mentions)
Iba 1, by Abcam (2 mentions)
Neuronal nuclei (neun), by Merck Group (2 mentions)
Cd11b, by Abcam (2 mentions)
Nf κb, by Cell Signaling Technology (2 mentions)
Easytaq dna polymerase, by Transgene (2 mentions)
Sodium thioglycolate, by Merck Group (2 mentions)
Brain derived neurotrophic factor (bdnf), by Abcam (2 mentions)
19 protocols using 4 6 diamidino 2 phenylindole dilactate dapi
1
Quantification of Inflammatory Markers in Cell Samples
2
Immunostaining of Brain and Colon Tissues
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Mice were trans-cardiacally perfused with 4% paraformaldehyde. Their brains and colons were post-fixed with 4% paraformaldehyde for 4 h, cytoprotected in 30% sucrose solution, freezed, cut using a cryostat (Leica, Nussloch, Germany), and immunostained according to the method of Jang et al. (11 (link)). Briefly, the sections were washed with phosphate buffered saline, blocked with normal serum, incubated with antibodies for Iba1 (1:200, Abcam), LPS (1:200, Millipore), NF-κB (1:100, Cell Signaling), CD11b (1:200, Abcam), CD11c (1:200, Abcam), and/or NeuN (1:200, Millipore) overnight, and treated with the secondary antibodies for 2 h. Secondary antibodies conjugated with with Alexa Fluor 488 (1:200, Invitrogen) or Alexa Fluor 594 (1:200, Invitrogen) were then treated to visualize. Nuclei were stained with 4′,6-diamidino-2-phenylindole, dilactate (DAPI, Sigma). Immunostained tissue slices were scanned with a confocal laser microscope.
3
Immunostaining of Brain and Colon Tissues
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Immunostaining was performed, as previously reported (Jang et al., 2018b (link),c ). Briefly, brain and colon tissue sections were washed with PBS, blocked with normal serum, incubated with antibodies for Iba1 (1:200, Abcam), LPS (1:200, Millipore), NF-κB (1:100, Cell Signaling), CD11b (1:200, Abcam), CD11c (1:200, Abcam), and/or NeuN (1:200, Millipore) antibodies overnight, and treated with the secondary antibodies for 2 h. Secondary antibodies conjugated with Alexa Fluor 488 (1:200, Invitrogen) or Alexa Fluor 594 (1:200, Invitrogen) were then treated to enable visualization. Nuclei were stained with 4’,6-diamidino-2-phenylindole, dilactate (DAPI, Sigma). Immunostained samples were scanned with a confocal laser microscope.
4
Antimony-Based Leishmania Treatment Assay
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Trivalent antimony (SbIII), amphotericin B (AmB), paromomycin, Triton X-100, paraformaldehyde, 4′,6-diamidino-2-phenylindole dilactate (DAPI), n-dodecyl-β-D-maltoside (DDM), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and phorbol 12-myristate 13-acetate (PMA) were obtained from Sigma-Aldrich (St. Louis, USA). Miltefosine was purchased from Æterna Zentaris (Frankfurt, Germany). Glucantime® was purchased from Sanofi-Aventis (Paris, France). L-glutamine and penicillin/streptomycin were obtained from Gibco. All chemicals were of the highest quality available.
5
Quantifying Cardiomyocyte Apoptosis via TUNEL Assay
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We detected apoptosis using the Dead End Fluorometric TUNEL System (Promega), according to the manufacturer's instructions. Four frozen hearts per group were sliced into 5 μm thick sections using a freezing microtome. Frozen sections were counterstained with mouse monoclonal α-sarcomeric actin antibody (1 : 75, Abcam), and Alexa Fluor 594 rabbit anti-mouse antibody (1 : 1000, Invitrogen) was applied as secondary antibody; cell nuclei were stained with 4′,6-Diamidino-2-Phenylindole, Dilactate (DAPI; Sigma). The stained sections were mounted and analyzed with a Fluoview 1000 confocal microscope (Olympus, Japan). We counted 6 random 400x fields and the number of TUNEL-positive cardiomyocyte nuclei was manually determined. The total number of nuclei (exhibited as DAPI-positive signals) was automatically calculated using Image-Pro Plus (version 5.0).
6
Microbial Immunology and Molecular Characterization
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Sodium thioglycolate, 4′,6-diamidino-2-phenylindole dilactate (DAPI), LPS, and RPMI 1640 were purchased from Sigma (St Louis, MO). A De Man, Rogosa, and Sharpe (MRS) medium, Sabouraud dextrose agar (SDA), and general anaerobic medium were purchased from BD (Franklin Lakes, NJ). An antibody for NF-κB was purchased from Cell Signaling Technology (Danvers, MA). Antibodies for CD11c, Iba1, and BDNF were purchased from Abcam (Cambridge, U.K.). Alexa Fluor 488 and Alexa Fluor 594 were purchased from Invitrogen (Carbsband, CA). Enzyme-linked immunosorbent assay (ELISA) kits for IFN-γ, TNF-α, and IL-10 were purchased from Ebioscience (Atlanta, GA). A QIAamp Fast DNA stool mini kit was purchased from Qiagen (Hilden, Germany). EasyTaq DNA polymerase and 100 bp plus II DNA ladder were purchased from TransGen Biotech (Beijing, China). TB Green® Premix Ex Taq™ II was purchased from Takara Bio (Shiga, Japan).
7
Cellular Visualization via Fluorescence Microscopy
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After three and 21 days of culture, the constructs were washed with PBS during 5 min., fixed with 10% formalin for one hour, and then washed again. Next, Phalloidin-TRITC (Sigma, USA) (1:80) was added and placed at room temperature during 1 h, protected from light. The constructs were counterstained with 4,6-Diamidino-2-phenylindole, Dilactate (DAPI, Sigma, USA) (1:5000) for 5 min. in the dark. Finally, they were visualized by Confocal Laser Scanning Microscope (TCS SP8, Leica, Mannheim, Germany).
8
Immunological and Neurological Assays
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A general anaerobic medium (GAM) and radioimmunoprecipitation assay (RIPA) lysis buffer were purchased from Nissui Pharm. Co., Ltd. (Tokyo, Japan) and Biosesang Inc. (Seongnam, Republic of Korea), respectively. Enzyme-linked immunosorbent assay (ELISA) kits for cytokines were purchased from Ebioscience (Atlanta, GA, USA). Antibodies for brain-derived neurotropic factor (BDNF) and NeuN were purchased from Millipore (Burlington, MA, USA). Antibody for NF-κB was purchased from Cell Signaling Technology (Beverly, MA, USA). Antibodies for Iba1 were purchased from Abcam (Cambridge, UK). 4′,6-Diamidino-2-phenylindole dilactate (DAPI) was purchased from Sigma (St. Louis, MO, USA). Enzyme-linked immunosorbent assay (ELISA) kits for TNF-α, IL-1β, and IL-10 were purchased from Ebioscience (Atlanta, GA, USA).
9
Apoptosis Induction Reagents Protocols
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AND, propidium iodide (PI), and 4,6-diamidino-2-phenylindole dilactate (DAPI) were purchased from Sigma. 3AB, Z-VAD, and DEVD were purchased from Biomol (Enzo Life Sciences Inc., NY, USA). PD98059 was purchased from Cell Signaling Technology Inc. (Beverly, MA, USA).
10
Immunocytochemical Staining of Neurons
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After 24 hours of culture, neurons were fixed at room temperature in 4% paraformaldehyde in PBS for 20 minutes, washed two times with PBS, and blocked using PBS, 10% of donkey serum, and 0.1% of Triton x100. Then, cells were washed twice and incubated with primary antibodies against rat monoclonalβ-tubulin III (1:1000, ab6160, Abcam) overnight at 4°C. Cells were washed three times again, 10 min each, and incubated with Donkey anti-rat Alexa 488 secondary antibodies (1:500, ab150153, Abcam) for 1 h, in dark at room temperature. Primary and secondary antibodies were diluted in PBS, 1% of donkey serum (GeneTex, Irvine, CA, USA) and 0.01% of Triton x100. Next, cultures were washed three times, 10 min each, and were counterstained with 4,6-Diamidino-2-Phenylindole, Dilactate (DAPI, 1:1000, Sigma). Glass slides were mounted in prolong mounting media, and preparations were cured overnight at +4°C protected from light until microscopy analysis.
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