Tmr dextran

Manufactured by Merck Group

Sourced in United States

TMR-dextran is a laboratory reagent that consists of a dextran polymer conjugated with the fluorescent dye tetramethylrhodamine (TMR). It is commonly used as a molecular probe or label in various biological research applications.

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Lab products found in correlation

Fitc dextran, by Merck Group (1 mentions) Prism 6, by GraphPad (1 mentions) Fluorescence plate reader, by Tecan (1 mentions) Oligomycin, by Merck Group (1 mentions) Pp242, by Selleck Chemicals (1 mentions) Bamhi, by New England Biolabs (1 mentions) Rapamycin, by Selleck Chemicals (1 mentions) P iκbα, by Cell Signaling Technology (1 mentions) C fos, by Cell Signaling Technology (1 mentions) Ova323 339, by InvivoGen (1 mentions) Bptes, by Selleck Chemicals (1 mentions) Ova257 264, by InvivoGen (1 mentions) Fluoro carbonyl cyanide phenylhydrazone (fccp), by Merck Group (1 mentions) Bms 303141, by Cayman Chemical (1 mentions) P acly, by Cell Signaling Technology (1 mentions) Bodipy, by Thermo Fisher Scientific (1 mentions) C myc, by Cell Signaling Technology (1 mentions) Mitotracker, by Thermo Fisher Scientific (1 mentions) Ku 0063794, by Selleck Chemicals (1 mentions) H3k9ac, by Cell Signaling Technology (1 mentions)

6 protocols using tmr dextran

Doxorubicin-Induced Epithelial Permeability

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bEnd5 cells were cultured on 24-well inserts to build a monolayer as described above, before being treated with DTX (5 ng/mL) or DMSO-control (0.25%) for 72 h. Afterwards, for permeability assay, fluorescent tracers of different sizes (0.45 kD LY (Sigma-Aldrich), 3 kD TXR dextran (Thermo Fisher Scientific, Dreieich, Germany), 20 kD TMR dextran (Sigma-Aldrich), 70 kD FITC dextran (Sigma-Aldrich)) were used as described previously [23 (link)] at the following timepoints: 1 h, 2 h, 3 h. Briefly, the tracer mix was added to the upper chamber and at each time-point media aliquots from both chambers were collected. Samples were read in a fluorescence plate reader (Tecan, Männedorf, Switzerland) at the corresponding tracer excitation/emission. Permeability was calculated as follows: bottom chamber fluorescence normalized to the apical chamber fluorescence with the ratio for the control condition set to 100% [25 (link)]. Statistical analysis was done using Prism 6.0 (GraphPad software), unpaired t-test.

Bernatz S., Ilina E.I., Devraj K., Harter P.N., Mueller K., Kleber S., Braun Y., Penski C., Renner C., Halder R., Jennewein L., Solbach C., Thorsen F., Pestalozzi B.C., Mischo A, & Mittelbronn M. (2019). Impact of Docetaxel on blood-brain barrier function and formation of breast cancer brain metastases. Journal of Experimental & Clinical Cancer Research : CR, 38, 434.

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Genetic Modification and Cell Signaling

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The H2-K^b was amplified from C57BL/6 splenocytes and cloned into a pCDH-GFP vector by enzymes EcoRI and BamHI (NEB). The Acaca and Acly shRNAs were cloned into a pLKO.1-GFP vector. LPS, OVA_257-264, OVA_323-339, and OVA were purchased from InvivoGen. TOFA, C75, 2-DG, 2-NBDG, etomoxir, and BMS-303141 were purchased from Cayman Chemical. Rapamycin, KU0063794, PP242, and BPTES were from Selleck. Mitotracker and BODIPY were purchased from Invitrogen. UK5099, 25-HC, TMR-dextran, oligomycin, fluoro-carbonyl cyanide phenylhydrazone (FCCP), rotenone, and antimycin A were from Sigma. Antibodies for TSC1 (#4906), TSC2 (#4308), c-Myc (#5605), c-Fos (#2250), p-S6 (#4858), S6 (#2317), IRF1 (#8478), IRF5 (#4950), ACC1 (#3662), H3K9ac (#9649), p-ACLY (#4331), ACLY (#4332), p-IκBα (#2859), p-IKKα/β (#2697), p65 (#8242), and CREB (#9197) were from Cell Signaling Technology. Antibodies against H3K27ac (ab4729), H3K27me3 (ab6002), H3K9me3 (ab8898), and IKKα/β (ab178870) were purchased from Abcam. Antibody for IκBα (SC371) was from Santa Cruz Biotechnology. Antibody against GAPDH (AP0063) was from Bioworld.

Shi L., Chen X., Zang A., Li T., Hu Y., Ma S., Lü M., Yin H., Wang H., Zhang X., Zhang B., Leng Q., Yang J, & Xiao H. (2019). TSC1/mTOR-controlled metabolic–epigenetic cross talk underpins DC control of CD8+ T-cell homeostasis. PLoS Biology, 17(8), e3000420.

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Quantifying Tumor Vasculature Extravasation

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Labeling flowing vasculature and sites of permeability was performed as previously described (10 (link)). Briefly, to quantify extravasation, high molecular weight 155 kDa TMR-dextran (cat# T1287, Sigma-Aldrich, Burlington, MA, USA) diluted in PBS was administered by tail vein i.v. one hour before the termination of the experiments. Anti-mouse CD31-biotin (cat# 13–0311-82, Thermo Fisher, Waltham, MA, USA) was administered by tail vein i.v. ten minutes before the end of the experiment to label flowing blood vessels. At time of sacrifice tumors were removed and fixed for 48 hours in 10% formalin in a volume ratio of tumor to formalin of 1:7 and made into paraffin blocks. Paraffin blocks of tumors were cut into 5 μm sections and immunofluorescence staining was performed. TMR-dextran is visualized using rabbit anti-TMR (A-6397; Life Technologies, Carlsbad, CA, USA).

Surve C.R., Duran C.L., Ye X., Chen X., Lin Y., Harney A.S., Wang Y., Sharma V.P., Stanley E.R., Cox D., McAuliffe J.C., Entenberg D., Oktay M.H, & Condeelis J.S. (2024). Signaling events at TMEM doorways provide potential targets for inhibiting breast cancer dissemination. bioRxiv.

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Multiphoton Imaging of Post-Stroke Vasculature

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For multiphoton imaging, we used an upright Zeiss LSM710 confocal microscope equipped with a Ti:Sa laser (Chameleon Vision II) from Coherent (Glasgow, Scotland) and 2 external photomultiplier detectors for red and green fluorescence.⁸⁸ (link) Anesthetized animals (1.5 % of isoflurane) were placed on a heating pad under the microscope. For visualization of the vasculature, 5 min prior to the imaging, the fluorescent tracer Tetramethylrhodaminisothiocyanat-Dextran (TMR-Dextran), 3000 Da MW (Sigma-Aldrich, St.Luis, MI, UA) was injected subcutaneously. The scanning was performed with Z-stack, 50-100 μm depth, laser (900 nm) power from 6-8% till 12-16% depending on the region of interest (ROI) depth. GAASP detector with LP˂570 nm filter for the GFP channel, LP˃570 nm for the TMR channel, and NDD detector SP˂485 nm for the bone visualization, all with master gain 600. Image size 1024x1024, 8 bit. Objective: W Plan-Apochromat 20x/1.0 DIC M27 75mm. For the series scanning, the laser power was 8-10%, 5 frames every 1 s. For each animal, 2-3 ROI was chosen which were imaged at baseline, 2, 24, and 72 hours post-stroke, or at the respective time point for naïve and sham-operated animals.

Kolabas Z.I., Kuemmerle L.B., Perneczky R., Förstera B., Ulukaya S., Ali M., Kapoor S., Bartos L.M., Büttner M., Caliskan O.S., Rong Z., Mai H., Höher L., Jeridi D., Molbay M., Khalin I., Deligiannis I.K., Negwer M., Roberts K., Simats A., Carofiglio O., Todorov M.I., Horvath I., Ozturk F., Hummel S., Biechele G., Zatcepin A., Unterrainer M., Gnörich J., Roodselaar J., Shrouder J., Khosravani P., Tast B., Richter L., Díaz-Marugán L., Kaltenecker D., Lux L., Chen Y., Zhao S., Rauchmann B.S., Sterr M., Kunze I., Stanic K., Kan V.W., Besson-Girard S., Katzdobler S., Palleis C., Schädler J., Paetzold J.C., Liebscher S., Hauser A.E., Gokce O., Lickert H., Steinke H., Benakis C., Braun C., Martinez-Jimenez C.P., Buerger K., Albert N.L., Höglinger G., Levin J., Haass C., Kopczak A., Dichgans M., Havla J., Kümpfel T., Kerschensteiner M., Schifferer M., Simons M., Liesz A., Krahmer N., Bayraktar O.A., Franzmeier N., Plesnila N., Erener S., Puelles V.G., Delbridge C., Bhatia H.S., Hellal F., Elsner M., Bechmann I., Ondruschka B., Brendel M., Theis F.J, & Erturk A. (2023). Distinct molecular profiles of skull bone marrow in health and neurological disorders. Cell, 186(17), 3706-3725.e29.

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Caco-2 Cell Permeability Assay

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TMR-dextran (MW: 65 -85 kDa) was purchased from Sigma-Aldrich (St. Louis, MO, USA). Quinidine sulfate dehydrate and Rh123 (MW: 381 Da) were purchased from Wako Pure Chemicals (Osaka, Japan). Calcein was purchased from Dojindo (Kumamoto, Japan). Caco-2 human colon carcinoma cell line was obtained from Riken BioResource Center (Ibaraki, Japan).

Sakuta Y., Takehara I., Tsunoda K.I, & Sato K. (2018). Development of a Microfluidic System Comprising Dialysis and Secretion Components for a Bioassay of Renal Clearance. Analytical sciences : the international journal of the Japan Society for Analytical Chemistry, 34(9).

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Tracking Phagosome-Lysosome Fusion in Macrophages

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For detection of phagosome-lysosome fusion, macrophages were preincubated with 1 mg/mL TMR dextran (10 kDa; Sigma-Aldrich) for 1 h in HEPES/MEM at 37°C. Cells were then washed with HEPES/MEM and infected with S. aureus as above. Cells were fixed and permeabilized as above. Lysosomes were stained with antibodies against anti-Lamp1 (Abcam) followed by incubation with FITCcoupled anti-rat antibodies. For observation of intracellular acidic organelles, macrophages were incubated with 60 nM LysoTracker Red DND-99 (Thermo Fisher) in MEM. Macrophages were then infected with S. aureus, washed, and analyzed with confocal microscopy, as described above.

Li C., Wu Y., Riehle A., Orian-Rousseau V., Zhang Y., Gulbins E, & Grassmé H. (2018). Regulation of Staphylococcus aureus Infection of Macrophages by CD44, Reactive Oxygen Species, and Acid Sphingomyelinase. Antioxidants & redox signaling, 28(10).

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