Cadmium acetate (CdAC2), GA, Lucifer yellow (LY) dilithium salt, rhodamine-labeled dextran (RD), Fluo-4/AM and Hoechst 33258 were purchased from Sigma-Aldrich (USA). An annexin V-FITC Fluorescence Microscopy Kit was obtained from BD Biosciences Pharmingen (USA). Dulbecco's modified Eagle's medium (DMEM) and fetal bovine serum were purchased from Gibco Laboratories (USA). Cx43, p-Cx43, Bax, Bcl-2, caspase-3, ERK, p-ERK, JNK, p-JNK, p-38, p-p38 and β-actin were purchased from Cell Signaling Technology (USA). All other chemicals and reagents used were of analytical grade and acquired locally.
Rhodamine labeled dextran
Manufactured by Merck Group
Sourced in United States
Rhodamine-labeled dextran is a fluorescent dye-labeled polysaccharide compound. It is designed for use as a tool in biological research, providing a means to visualize and track the movement and distribution of dextran in various experimental systems.
Automatically generated - may contain errors
Lab products found in correlation
Hoechst 33258, by Merck Group (1 mentions)
Bcl2 associated x (bax), by Cell Signaling Technology (1 mentions)
P cx43, by Cell Signaling Technology (1 mentions)
Annexin 5 fitc fluorescence microscopy kit, by BD (1 mentions)
Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (1 mentions)
P erk, by Cell Signaling Technology (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Caspase 3, by Cell Signaling Technology (1 mentions)
P jnk, by Cell Signaling Technology (1 mentions)
Bcl 2, by Cell Signaling Technology (1 mentions)
β actin, by Cell Signaling Technology (1 mentions)
Fluo 4 am, by Merck Group (1 mentions)
Infinite 200 pro, by Tecan (1 mentions)
Fluostar omega plate reader, by BMG Labtech (1 mentions)
4 protocols using rhodamine labeled dextran
1
Apoptosis Signaling Pathway Analysis
2
Dextran-Aided Bleb Dynamics Tracking
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Cells expressing EGFP or Emerald-tagged protein were plated on uncoated 6-well glass bottom plates (In Vitro Scientific) in media containing 0.1 mg/ml rhodamine-labeled dextran (MW = 70 kDa, Sigma Aldrich). Time of arrival along the bleb perimeter was determined to be when EGFP or Emerald fluorescence was above background. Maximal protrusion of the bleb membrane as judged by displacement of rhodamine-dextran (negative stain) was set as time 0.
3
Dextran-Rhodamine Release from Chitosan Capsules
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A high molecular weight (~150,000 g/mol) rhodamine-labeled dextran (Sigma-Aldrich) was used as a model protein. We mixed 30 μL of dextran–rhodamine (20 mg/mL) with 300 μL chitosan and generated the capsules. The chitosan capsules were first washed by PBS for three times and then cultured in 3 mL M9 medium. As control, same amount of plain capsules were cultured in 3 mL M9 media. In total, 100 μL of the culturing medium was taken at different time points and the fluorescence signal from the medium was measured by a platereader (Infinite 200pro, TECAN). Experiments were done in triplicate and measurements were background corrected. Unless mentioned otherwise, plots in main figures and supplementary figures were generated by Excel (2013).
4
Gastrointestinal Transit Assay in Mice
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Gastrointestinal transit was analyzed as previously described [5] . Briefly, mice were fasted overnight and were allowed free access to water. In the morning mice were given 100 μl of 25 mg/ml rhodamine labeled dextran (Sigma-Aldrich) solution by gavage. 40 min after gavage, the mice were sacrificed and the gastrointestinal tract, from stomach to cecum, was removed and placed in cold saline. The small intestine was divided into 10 equal sections and each segment, in addition to the stomach, was flushed with 2 ml of saline. The flushed contents were centrifuged at 500 rpm for 10 min and 200 μl of the supernatant from each section was placed in a 96 well plate. The quantification of the fluorescent signal in the supernatant from each segment was determined utilizing a multi-well fluorescence plate reader (FLUOstar Omega plate reader; BMG Labtech; excitation 545 nm and emission 590 nm). The distribution of the fluorescent signal in the intestinal segments was used to calculate the geometric center of fluorescence (GCF). GCF was determined by calculating the fraction of fluorescence per segment multiplied by the segment number (1-10) and adding all segments together. GCF can range from 1 to 10 with a higher number indicating a faster intestinal transit time.
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