65-2860 Fixable Viability Dye eFluor506/780 S ample P ck - Table 1: Table of Fixable Viability Dyes General Notes Best practices when using Fixable Viability Dyes 1. About eFluor 506. eFluor 506 (eF506) is a green-emitting organic dye that can be excited by the 405 nm laser and collected using a 510/50 nm bandpass filter. No. 5. with whole blood from healthy donors. Cells were then xed and permeabilized (eBioscience, San Diego, CA, USA) over-night prior to incubation with mAb specic to Ki-67 (Ki-67). They should be protected from light and eFluor 615 . After washing, cells were incubated for 15 min with Fixable Viability Dye eFluor 506 (Thermo Fisher Scientific), washed and stained with CD3-PerCP-Vio700, rat (Miltenyi Biotec), goat F(ab') 2 anti rat IgG:FITC (Bio-Rad Laboratories) and R-Phycoerythrin AffiniPure F(ab') 2 The cells were then washed and incubated with primary anti- bodies against surface markers or matched isotype con- trol for 20min at 4C. 65-0866) -off the violet laser Fixable Viability Dye eFluor 520 (cat. Azide To make 10 mL of Azide, dissolve 2 g Azide in 10 mL of ultra-pure water (final concentration 20% w/v). eFluor 570 . eFluor 5506 has an excitation and emission peak at 405 nm and an emission peak at 506 nm, and is a suitable alternative to AmCyan or BD Horizon V500 for flow cytometry. Fixable Viability Dye eFluor 506 is a viability dye that can be used to irreversibly label dead cells prior to cryopreservation, fixation and/or permeabilization procedures. Store at 80C in 50 L aliquots for up to 6 months. (FITC, R1-2) and 7 integrin (PE, FIB27, all Biolegend), fixable Viability Dye efluor 506 (eBioscience). They should be protected from light and We wanted to quantify the effects of various treatments on cell death/membrane permeability as compared to controls under different conditions. FVD are supplied as a pre-diluted solutions prepared in high-quality, anhydrous DMSO. V7 BV510, eFluor 506, Amethyst Orange, VioGreen, Zombie Aqua, OC515, V500, LIVE/DEAD Aqua, Krome Orange, Ghost Dye Violet 510 & 540, Fixable Viability Stain 510 V8 cFluor V547, BV570, Pacific Orange, LSSmOrange, LIVE/DEAD Yellow, Live-or-Dye 405/545 V10 cFluor V610, BV605, Qdot 605, Super Bright 600, Zombie Yellow, Fixable Viability Stain 575V 535/45. 65-2860 Fixable Viability Dye eFluor506/780 S ample P ck - Table 1: Table of Fixable Viability Dyes General Notes Best practices when using Fixable Viability Dyes 1. Alternatively if the 65-0868) Note: The antibodies used for surface staining can be added after BrdU and Fixable Viability Dye labeling (but before fixation). Unlike 7-AAD and PI, cells labeled with Fixable Viability Dyes can be washe SKU: 65-0866-18 + Life Technologies (Thermo Fisher) Antibiotic-Antimycotic (100X) $ 36.52. Invitrogen Alexa Fluor 555 [Optional] Fixable Viability Dyes eFluor 450, 506, 660 and 780 (eBioscience Cat. Fixable Viability Dye eFluor 506 , eBioscience Supplier: eBioscience Fixable Viability Dye eFluor 506 is a viability dye that can be used to irreversibly label dead cells prior to cryopreservation, fixation and/or permeabilization procedures. Fixable Viability Dye eFluor 506 is supplied as a pre-diluted solution prepared in high-quality, anhydrous DMSO It should be protected from light and moisture Store at less than or equal to -70C with dessicant, it may be freeze-thawed up to 20 times Allow vial to equilibrate to room temperature before opening Specifications - Fixable Viability Dye eFluor 506 (Cat. To help with pinpointing exact data, the analyzer has the capacity to zoom in, allowing you easily visualize spectra intersection points with laser lines, filter sets and other spectra. 65-0867) and eFluor 455UV (Thermo Fisher Cat. 695/40. 65-0866) or Fixable Viability Dye eFluor 780 (Cat. Fixable Viability Dye eFluor 506 405 506 ThermoFisher 65-0866-14 Dead Fixable Permeable Amino Ghost Dye Violet 450 405 450 Tonbo 13-0863-T100 Dead Fixable Impermeant Amino Ghost Dye Violet 510 405 510 Tonbo 13-0870-T100 Dead Fixable Impermeant Amino Allow vial of FVD to equilibrate to room temperature before opening. [Optional] Fixable Viability Dyes eFluor 450, 506, 660 and 780 (eBioscience Cat. For detection of cytokine production with non-specic stimulation, obtained PBMCs were stimulated for 4.5 h at Zoom in, zoom out, and pan. eFluor 520 . Fixable Aqua Dead cell stain, Fixable Viability eFluor 506, Krome Orange, Pacific Orange, Qdot 525, VioGreen: 560/20* 550LP: Qdot 565, Pacific Orange, Krome Orange : 585/15* 575LP: Brilliant . Can be used to irreversibly label dead cells prior to cryopreservation, fixation and/or permeabilization procedures. Can be used to irreversibly label dead cells prior to cryopreservation, fixation and/or permeabilization procedures. In these experiments, we treated mice in vivo with different inhalants/particulates and used this FVD eFluor 506 to determine BAL cell viability as a part of a larger flow cytometric panel. They should be protected from light and - Fixable Viability Dye eFluor 506 (Cat. 65-0865), eFluor 506 (Thermo Fisher Cat. However, in late Store at 4C for up to 1 year. Fixable viability dye eFluor 506: Thermo Fisher (CA, USA) 65-0866-14: Critical Commercial Assays; Mouse tumor dissociation kit: Myltenyi Biotech (Germany) 130-096-730: Chromium Single Cell 5 Library and Gel Bead Kit: 10x Genomics (CA, USA) 1000092: Deposited Data; Study data are deposited in GEO: Zombie Aqua Fixable Viability Kit is composed of lyophilized Zombie Aqua dye and anhydrous DMSO. Store at 80C in 50 L aliquots for up to 6 months. No. For surface staining of PBMC or isolated T cells, antibodies were incubated for 30 min at 4 C and washed twice with phosphate buffered saline (PBS). Wash cells one time in sodium azide- and protein-free Dulbecco's Phosphate Buffered Saline (1X DPBS). Fixable Viability dye eFluor-506 as described above; co-stained with anti-CD3, anti-CD4, anti-CD28, anti-CX3CR1, anti-CD49d and anti-CD11b antibodies; and fixed and permeabilised as already described, followed by intracellular staining with anti-perforin and anti-granzyme B antibodies. 65-0866) . For reconstitution, pre-warm the kit to room temperature; add 100 l of DMSO to one vial of Zombie Aqua dye and mix until fully dissolved. In early-stage apoptosis, the plasma membrane excludes viability dyes such as propidium iodide (PI), 7-AAD, as well as Fixable Viability Dyes (FVD) such as eFluor 660, eFluor 506 or eFluor 780. They should be protected from light and no. 610/20. It can be excited using a 405 nm laser paired with a 525/50 nm bandpass filter, a configuration that can be found, for example, in the BD FACSymphony A5. Please take this . Store at 80C in 50 L aliquots for up to 6 months. no. No. We also recommend the use of the Fixable Viability Dyes to help eliminate dead cells during the analysis. 88-8824) n Flow Cytometry Staining Buffer (cat. 1. GFP . To make 10 mL of Azide, dissolve 2 g Azide in 10 mL of ultra-pure water (final concentration 20% w/v). Fixable Viability Dyes eFluor 506 (405 nm) 506 nm 510/50 AmCyan Fixable Viability Dye eFluor 570 . No. Resuspend cells at 1-10x10^6 cells/ml in sodium azide- and protein-free 1X DPBS. 65-2860 Fixable Viability Dye eFluor506/780 S ample P ck - Table 1: Table of Fixable Viability Dyes General Notes Best practices when using Fixable Viability Dyes 1. Fixable viability dye Prepare a working solution of the Fixable Viability Dye eFluor 506 by adding 450mLof13 D-PBS to the vial (nal concentration 1:10). As fixation and permeabilization will impact the brightness of eFluor nanocrystals, we . Invitrogen Alexa Fluor 488 . Transfer the blood sample to a 15 mL tube containing 5 mL of ACK lysis buffer, close tube, vortex shortly (5 s) and incubate at RT for 3 min. Unlike 7-AAD and propidium iodide, cells labeled with Fixable Viability Dyes can be washed, fixed, permabilized, and stained for intracellular antigens without any loss of Fixable Viability Dye eFluor 506 and 780 are viability dyes that can be used to irreversibly label dead cells prior to cryopreservation, fixation and/or permeabilization procedures. Fixable Viability Stain 700 . Samples were fixed with FoxP3/Transcription Factor Staining Buffer Set (eBioscience). R4: cFluor R720, Alexa Fluor 680, Alexa Fluor 700, APC-Alexa Fluor 700, APC-R700, Fixable Viability Stain 700, Ghost Dye Red 710, R718, redFluor710. Put the tubes back on ice and add 8 mL cold T cell culture medium to stop red blood cells lysis. Unlike 7-AAD and propidium iodide, cells labeled with Fixable Viability Dyes can be washed, fixed, permabilized, and stained for intracellular antigens without any loss of staining intensity of the dead cells. EYFP . Ghost Dye UV 450 . FITC . Ghost Dye Violet 510 . (A) Forward and side scatterplot of viable . 4. . PE-eFluor 610 . A custom collection of spectra representing the microscope Nikon Eclipse TiE Inverted Microscope (Fluorescent Widefield) on FPbase. 65-0865) Buffers and solution preparation Prepare fresh Fixation/Permeabilization working solution by diluting the Fixation/Permeabilization Concentrate (1 part) with Fixable viability dye Prepare a working solution of the Fixable Viability Dye eFluor 506 by adding 450 L of 1 D-PBS to the vial (final concentration 1:10). 3.5 Intracellular Staining. Fixable Viability Dyes eFluor 455UV, eFluor 450, eFluor 506, eFluor 520, eFluor 660 and eFluor 780 are permanent dyes suitable for UV, violet, blue and red lasers that will not wash out of cells. To zoom in, double click your mouse over the area that you wish to zoom, or roll the mouse wheel forward. 65-0863, 65-0866, 65-0864, 65-0865) Experimental Procedure . Store at -70C with dessicant. Cell viability during flow cytometric analyses was determined using fixable viability dye eFluor 506 (eBioscience, San Diego, CA, USA). . CellVue Plum, Cy5, Cy5.5, DRAQ5, DRAQ7, Dye Cycle Ruby, Fixable far red dead stain, Fixable Viability eFluor 660, Fx Far Red Cell Stain, SYTOX Red . Dead cell populations have high fluorescent intensity, allowing for easy exclusion, thereby improving data quality. PDF | Flow cytometry immunophenotyping is a sensitive technique allowing rapid characterization of single cells within heterogeneous populations, but it. (AmCyan channelfixable viability stain eFluor 506, V450violet proliferation dye 450). 00-6990), or Fixable Viability Dyes eFluor 450, 506, 660 and 780 (Cat. We also recommend the use of the Fixable Viability Dyes to help in the analysis. cFluor R668, Alexa Fluor 647, eFluor 660, Fixable Viability Stain 660, Janelia Fluor 646, LIVE/DEAD Far Red, Live-or-Dye 640/662, Northern Lights 637, SYTOX Deep Red, Vio 667, VioBright 667. No. no. a. eFluor 780 (Thermo Fisher Cat. Ghost Dye Violet 540 . 3. No. If cells were not prelabeled with a Fixable Viability Dye in Step 1, stain samples with a viability dye, according to the Viability Staining Protocol in our Best . Ghost Dye Blue 516 . Unlike 7-AAD and PI, cells labeled with Fixable Viability Dyes can be washed, fixed, permabilized and stained for intracellular antigens without loss of staining intensity. . Fixable Viability Dye eFluor 506 can be excited by the violet (405 nm) laser line and has a peak emission of 506 nm that can be detected using a 510/50 band pass filter (equivalent to AmCyan) For compensation, it is recommended to use a sample of the cells of interest stained with the Fixable Viability Dye; - Fixable Viability Dye eFluor 780 (Cat. SKU: 15596018 + Life Technologies (Thermo Fisher) ProLong Gold Antifade Mountant eFluor 780 . Storage & Handling 65-0864) -off the red laser 65-0866), eFluor 520 (Thermo Fisher Cat. No. 2. 585/40. Thus, using Fixable Viability Dyes allows dead cells to be excluded from analysis when intracellular targets are being studied. 65-0865) Buffers and solution preparation Prepare fresh Foxp3 Fixation/Permeabilization working solution by diluting the Fixation/Permeabilization Concentrate (1 part) In some cases, the iframe may appear truncated if inserted into a div with smaller height. 65-0867) -off the blue laser Fixable Viability Dye eFluor 660 (cat. Viability dyes (compatible with fixation) Viability dyes (non-fixed cells) DNA content/cell cycle dyes (live cells) DNA content/cell cycle dyes (fixed cells) Apoptosis dyes Cell proliferation dyes ROS detection dyes Phagocytosis dyes Fluorescent proteins Other dyes Violet (405 nm) 440/50 VL1 Alexa Fluor 405 Pacific Blue LIVE/DEAD Fixable . FVD are supplied as a pre-diluted solutions prepared in high-quality, anhydrous DMSO. markers (Antibodies and Staining Reagents below), then stained with the fixable viability dye eFluor 506 (eBioscience) in HBSS for 25 minutes. Ghost Dye Red 780 . Item Fixable Viability Dye eFluor 506 500 tests; Company Thermo Fisher Scientific; Catalog Number 65-0866-18; This product is no longer available on Biocompare. No. b. It should be protected from light and moisture. # 650866) was used for cell viability in parallel with antibody staining, following the manufacturer's instructions. Store at 4C for up to 1 year. Fixable Viability Dye eFluor 506 is a fluorescent compound with an excitation peak at 409 nm and an emission peak at 506 nm, giving it a fairly large Stokes' Shift of 97nm. Fixable Viability Dye eFluor 450 (cat. Prepare a working solution of the Fixable Viability Dye eFluor 506 by adding 450 L of 1 D-PBS to the vial (final concentration 1:10). 00-4222) BV510, eFluor 506, Amethyst Orange, VioGreen, Zombie Aqua, OC515, V500, LIVE/DEAD Aqua, Krome Orange, Ghost Dye Violet 510 & 540, Fixable Viability Stain 510 V8 cFluor V547, BV570, Pacific Orange, LSSmOrange, LIVE/DEAD Yellow, Live-or-Dye 405/545 No. . 65-0863, 65-0866, 65-0864, 65-0865) Directly conjugated antibodies specific for . c. Centrifuge the sample at 400 g for 4 min at 4C. No. Fixable Viability Dye eFluor 506 can be excited by the violet (405nm) laser line and has a peak emission of 506nm that can be detected using a 510/50 band pass filter (equivalent to AmCyan) For compensation, it is recommended to use a sample of the cells of interest stained with the Fixable Viability Dye; n Fixable Viability Dyes eFluor 455UV, 450, 506, 520, 660 and 780 (cat. Cell samples were taken prior and post-magnetic selection followed by staining with a fixable viability dye (eFluor 506, eBiosciences), anti-CD8-APC (clone 53-6.7, BioLegend), anti-CD4-FITC (clone GK1.5, BD Pharmingen), and anti-CD19-PerCPCy5.5 (clone 1D3, BD Pharmingen) to check sample purity. FVDs are supplied as pre-diluted solutions prepared in highquality, anhydrous DMSO. Unlike 7-AAD and PI, cells labeled with Fixable Viability Dyes can be washed, fixed, permabilized and stained for intracellular antigens without loss of staining intensity. Store at 4C for up to 1 year. Fixable viability dye Prepare a working solution of the Fixable Viability Dye eFluor 506 by adding 450 L of 1 D-PBS to the vial (final concentration 1:10). These cells will stain with Annexin V but not with viability dyes, thus distinguishing cells in early apoptosis. VivaFix Cell Viability Dyes covalently bind to free primary amines which in live cells are present on the cell surface . Allow vial to equilibrate to room temperature before opening. Biocompare is the leading resource for up-to-date product information, product reviews, and new technologies for life scientists. Stained cells were measured on LSRII . Measurement of soluble markers of inflammation As fixation and permeabilization will impact the brightness of eFluor nanocrystals, we . - Fixable Viability Dye eFluor 780 (Cat. Prepare cells for flow cytometric staining using sodium azide-free buffers. Fixable Viability Dyes VivaFix Cell Viability Assays enable researchers to accurately discriminate between live and dead cells in unfixed samples by cell staining prior to flow cytometry, cell sorting and microscopy. PE-eFluor 610 . PerCP-eFluor 710 . eFluor 450, and eFluor 506 channels, but other channels (up to approximately 650 nm) may also be affected and depends on the cell type, laser line, and instrument settings. Wash the cells three times with PBS 1 at 350 g for 5-7 min at 4 C. Annexin conjugated to APC, eFluor 450, FITC, PE or PerCP-eFluor 710 Fixable Viability Dye eFluor 660 (Cat. Fixable Viability Stain 780 labeling of cells 1. In early-stage apoptosis, the plasma membrane excludes viability dyes such as propidium iodide (PI), 7-AAD, or Fixable Viability Dyes (FVD) such as eFluor 660, eFluor 506 or eFluor 780. . 100 tests = 1 vial of Zombie Aqua + DMSO, 500 tests = 5 vials of Zombie Aqua + DMSO. SVF cell suspension was stained with different CD markers and dead cells were excluded by using Ebioscience eFluor 506 fixable viability dye labelling. Unlike 7-AAD and propidium iodide, cells labeled with Fixable Viability Dyes can be washed, fixed, permabilized, and stained for intracellular antigens without . | Find, read and cite all the research . Ghost Dye Violet 450 . Fixable Viability Stain 660 . 65-0863, 65-0866, 65-0864, 65-0865) They should be protected from light and moisture. eFluor 570 . Fixable Viability Dye eFluor 506/780 Sample Pack; eBioscience: Can be used to irreversibly label dead cells prior to cryopreservation, fixation and/or permeabilization procedures. 5. Another group of trackers is represented by the fixable viability dyes, enabling the post-fixation discrimination of dead cells, which lost plasma membrane integrity before fixation . Fixable Viability Dye eFluor 506 (eBioscience, Cat. No. 4.4. 11. Fixable Aqua Dead cell stain, Fixable Viability eFluor 506, Krome Orange, Pacific Orange, Qdot 525, VioGreen V 610/20 BV605, eFluor605NC , Fixable Yellow Dead Cell Stain, Pacific Orange, Qdot 605 . 65-0864), Fixable Viability Dye eFluor 506 (Cat. To exclude dead cells from analysis, cells were incubated with Fixable Viability Dye eFluor 506 (eBioscience, San Diego, CA, USA) for 30min at 4C in the dark. V450 staining allows to track thymocyte proliferation and . 65-0866) . The staining was carried out using 50 L peripheral blood mixed with the reagents, followed by 10-15 min incu- EGFP . It may be freeze-thawed up to 20 times. (EH12.2H7), ICOS (C398.4A), and Fixable Viability Dye eFluorTM 506 at 4 C for 30 min. PerCP-eFluor 710 . eFluor 660 . APC, Alexa Fluor 647, Cell Proliferation Dye eFluor 670, CellVue Maroon, CellVue Plum, Cy5, Cy5.5, DRAQ5, DRAQ7, Dylight 649, Dye Cycle Ruby, Fixable . Flow Cytometry. Analyses were performed on LSR Fortessa (BD) and MACSQuant . These cells will stain with Annexin V but not a viability dye, thus distinguishing cells in early apoptosis. 65-0863) -off the violet laser Fixable Viability Dye eFluor 506 (cat. Fixable Viability Dye eFluor 506 is supplied as a pre-diluted solution prepared in high-quality, anhydrous DMSO. Azide To make 10 mL of Azide, dissolve 2 g Azide in 10 mL of ultra-pure water (final concentration 20% w/v). (DATK32, BioXCell) or IgG isotype (Sigma) as well as Hoechst dye and TexasRed Dextran (Life . No. Fixable Viability Dye eFluor 506 can be excited by the violet (405 nm) laser line and has a peak emission of 506 nm that can be detected using a 510/50 band pass filter (equivalent to AmCyan) For compensation, it is recommended to use a sample of the cells of interest stained with the Fixable Viability Dye; Store at less than or equal to -70C with dessicant. 65-0868, 65-0863, 65-0866, 65-0867, 65-0864, 65-0865) n Directly conjugated antibodies specific for intracellular proteins n Intracellular Fixation and Permeabilization Buffer Set (cat. eFluor 506 . 506: Fixable Viability Dye eFluor 520 d: 488: 488 nm: 522: Fixable Viability Dye 3Fluor 660 d: 633: 633 nm, 640 nm: 660: Fixable Viability Dye eFluor 780 d: 633: 633 nm, 640 nm: 780: a Thermo Fisher Life Technologies (all names are trademarked) b BioLegend (all names are trademarked) Fixable Viability Stain 780 . Note: pay attention to the height/width of the parent element. APC, Alexa Fluor 647, Cell Proliferation Dye eFluor 670, CellVue Maroon, CellVue Plum, Cy5, Cy5.5, DRAQ5, DRAQ7, Dye Cycle Ruby, Fixable far red dead stain, Fixable Viability eFluor 660, Fx Far Red Cell Stain, SYTOX Red, TO-PRO-3 R 730/45 APC-Cy5.5, Alexa Fluor 700, CellVue Burgundy , DRAQ7, Dye Cycle Ruby R 780/60 BD Bioscience) and a viability dye (eFluor 506 . No. Ghost Dye Red 710 . They may be freeze-thawed up to 20 times. 5. After the last wash, vacuum the supernatant and resuspend the cells in PBS 1 containing Fixable Viability Dye eFluor 506 (see Note 8) and incubate for 30 min on ice protected from light. d. Addition of a Fixable Viability Dye is recommended to ensure analysis is restricted to those cells that were alive at the start of the protocol. Propidium Iodide Staining Solution (Cat. 65-0865) : Fixable Viability Dye eFluor 450AnnexinV FVD are supplied as a pre-diluted solutions prepared in high-quality, anhydrous DMSO. No. No. R5 . eBioscience Fixable Viability Dye eFluor 506 $ 345.00. SKU: 15240062 + Life Technologies (Thermo Fisher) TRIzol Reagent $ 345.39. Store at 80 Cin50mL aliquots for up to 6 . 65-2860 Fixable Viability Dye eFluor506/780 S ample P ck - Table 1: Table of Fixable Viability Dyes General Notes Best practices when using Fixable Viability Dyes 1. FVD are supplied as a pre-diluted solutions prepared in high-quality, anhydrous DMSO. . Since NADH exhibits a characteristic autofluorescent peak at 470nm after excitation by a UV laser (27), mitochondrial NADH was calculated as a ratio of basal Reported Application
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