zombie aqua biolegend

2022년 1월 1일
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biolegend.com 11 Annexin V ZERO BIAS - scores, article reviews, protocol conditions and more Bioz Stars score: 99/100, based on 1 PubMed citations. Our affordable spectral flow cytometry instruments make complex applications possible. Newly developed pseudogout arthritis after therapy with ... Zombie Violet dye is excited by the Violet laser (405 nm) and has fluorescence emission maximum at 423 nm. Manufacturer: Biolegend 423102. Zombie UV™ is a polar water soluble dye, providing violet fluorescence, making it suitable for multi-color detection. Biolegend. A single-cell suspension was stained with Zombie Aqua Fixable Viability Kit (Biolegend), resuspended in FACS buffer (1% bovine serum albumin in saline), and incubated in block buffer prior to surface staining with anti-CD11b antibody (Novus Biologicals, #NB110-89474PECY7, polyclonal) for macrophages, monocytes, and granulocytes. If using in a multi-color panel design, filter optimization may be required depending on other fluorophores used. Cells were then fixed, The following dyes and antibodies were used to stain the cells: Zombie Aqua (Biolegend, San Diego, CA), CD45.2-PE/Dazzle 594 (Biolegend), CD45.1-Qdot 605 (Biolegend), IL17-Alexa 700 (Biolegend) and TNFα-Alexa 488 (Biolegend). more info or order : BioLegend product webpage. Thus, it can be used to assess live vs. dead status of mammalian cells. Zombie Aqua dye is excited by the violet laser and has a maximum emission of 516 nm. Live/dead cell staining was done together with the antibody incubation (Zombie NIR or Zombie Aqua, BioLegend, 1:500). BioLegend percp anti mouse cd45. mented with Zombie Aqua™ Fixable Viability Kit (final conc. Zombie Aqua™ dye is excited by the violet laser and has a maximum emission of 516 nm. The most commonly used surface markers for the identification of microglia include CD45 (low-intermediate expression), CD11b, Tmem119, and P2RY12. All samples examined by flow cytometry have been treated with Fc block (BD, 564219) prior to staining. Alternatively, autologous HBsAg-pulsed and HBcAg-pulsed Price from $9.99 to $1999.99. In cases where cell fixation is required, we now introduce fixable Zombie Aqua™, Zombie Green™, Zombie NIR™, Zombie Red™, Zombie Violet™, Zombie UV™, and Zombie Yellow™. It is critical to understand the degree of cell death in any flow cytometry assay and exclude those cells from the analysis. Cells were subsequently washed with staining buffer (PBS supplemented with 2% FCS) and stained with fluorescently conjugated antibody anti-CD8-AF647 (clone RPA-T8; BioLegend) for 20 minutes at 4°C. Our mission is to accelerate research and discovery by providing the highest quality products at an outstanding value, along with superior customer service and technical support. The NEW Biolegend Zombie dyes are a cheaper alternative to Invitrogen's fixable dyes. Buy from Supplier. Following fluorochrome-conjugated antibodies were used: CD42b PerCP-Cy5.5 (clone HIP1) and CD14 FITC (clone M5E2) from BioLegend, San Diego, CA. For live/dead staining, Zombie Aqua (BioLegend, San Diego, California) was included. (eBioscience, BD Biosciences, Biolegend) in the presence of anti-CD16/32 (2.4G2). Cells were cultured for 72 h in full medium in flat-bottom 96-well plates with stimulants as in cell survival assays, before harvesting and analysis on a flow cytometer. Clinical response rates may be improved by more immunogenic vaccines that COVID-19 Update: Learn about our efforts to help battle coronavirus. Anti-CD19-APC/Cy7 (clone HIB19, Biolegend) and anti-CD3-AlexaFluor700 (clone UCHT1, Biolegend) were added to exclude B cells and T cells, respectively. Flow cytometric measurements were performed on a BD LSRFortessa™ cytometer. Dilute Zombie Aqua Fixable Viability Kit (BioLegend) at a dilution of 1:200 in PBS. Cells alone without Zombie Aqua staining are indicated by the dashed histograms. percp anti mouse cd45 ( BioLegend ) 96. Single‐cell suspensions of blood (~3 × 10 6 -5 × 10 6 total cells) were incubated in PBS for 15 min at 4°C in the dark with the Zombie UV™ or Zombie Aqua™ Fixable Viability Kit (BioLegend, London, UK) as appropriate and immunoglobulin (Ig)G from human serum (Sigma Aldrich, Gillingham, UK). Zombie Violet™ is an amine-reactive fluorescent dye that is non-permeant to live cells but permeant to the cells with compromised membranes. KLRG1+ and KLRG1- CD56dim NK cells were sorted to a purity of ≥98% with the BD FACSAria III cell sorter (BD Biosciences). We analyzed data by using FACS Diva version 8.0.1 (BD Biosciences) and determined percentage viability by using Zombie Aqua (Biolegend) live-dead dye. BioLegend provides an extensive array of research tools . Item Zombie Aqua (TM) Fixable Viability Kit. Zombie Aqua™ is an amine-reactive fluorescent dye that is non-permeant to live cells but permeant to cells with compromised membranes. Thus, it can be used to assess live vs. dead status of mammalian cells. Cells were washed in PBS and stained for a . Percp Anti Mouse Cd45, supplied by BioLegend, used in various techniques. ZERO BIAS - scores, article reviews, protocol conditions and more 4/20/2020 Blue Zombies Gregor Werba Easy to use viability dye with emission spectrum that integrates nicely with Brilliant Violet dyes. BioLegend develops world-class, cutting-edge antibodies and reagents for biomedical research, manufactured in our state-of-the-art facilities in San Diego, CA. Bioz Stars score: 96/100, based on 1 PubMed citations. CANCER RESEARCH | TUMOR BIOLOGYAND IMMUNOLOGY Lymphatic PD-L1 Expression Restricts Tumor-Speciﬁc CD8þ T-cell Responses Nikola Cousin, Stefan Cap, Manuel Dihr, Carlotta Tacconi, Michael Detmar, and Lothar C. Dieterich Cellular viability was estimated by 7-AAD incorporation (BioLegend). LIVE/DEAD Zombie Aqua ™ (BioLegend®), anti-CD16 BUV395 (3G8, BD Horizon ™), anti-CD19 PE (HIB19, BioLe-gend®), anti-CD3 PerCP/Cyanine 5.5 (SK7, BioLegend®), anti-HLA-DR Alexa Fluor® 488 (L243, BioLegend®), anti-CD123 PE (6H6, BioLegend®), anti-CD11c PE-Cy7 (Bu15, BioLegend®), anti-CD14 Alexa Fluor® 700 (MSE2, Bio- It is critical to understand the degree of cell death in any flow cytometry assay and exclude those cells from the analysis. were stained with a viability dye (Zombie Aqua, Biolegend), Fc blocked (BD Biosciences) and incubated with a surface marker antibodies (CD3, CD4, CD8). ELISA Mouse flow . more info or order : BioLegend product webpage. Product Details All samples examined by flow cytometry have been treated with Fc block (BD, 564219) prior to staining. After stained with zombie aqua and the surface markers, splenocytes were fixed in fixation buffer (Biolegend) for 20 min at room temperature, permeabilized with intracellular staining wash buffer, blocked with anti-CD16/32, and then stained with anti-IFN-γ, granzyme B, and perforin. Zombie NIR™ is a polar water soluble dye, providing red fluorescence, making it suitable for multi-color detection. 28. 3 for 30min at 4°C. Bioz Stars score: 99/100, based on 1 PubMed citations. Single cell suspensions were incubated with purified anti-CD16/32 and UV-excitable LIVE/DEAD (UVLD) Blue (Invitrogen) or Zombie Aqua (Biolegend) and subsequently stained with a cocktail of fluorescently-labelled antibodies. If using in a multi-color panel design, filter optimization may be required depending on other fluorophores used. BioLegend products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products, reverse engineer Immunohistochemistry Fresh tumor samples were harvested after euthanasia, fixed in 4% PFA overnight, and preserved in 70% ethanol For intracellular cyto-kine staining, single cell suspensions of spleens or tumors Flow cytometry To assess surface expression of CD16 and NKG2D, cells were washed in 1% FBS/PBS (washing buffer) and blocked with 2% AB human serum/PBS (Sigma-Aldrich) for 10 min at 4°C and stained with viability dye (Zombie Aqua; BioLegend), BV421-labeled anti-CD56 mAb (clone HCD56; BioLegend), Alexa Fluor 647-labeled anti-CD16 mAb (clone . BioLegend provides DNA dyes, Propidium Iodide and 7-AAD, that enter and stain dead cells, but are impermeable to live cells for rapid, cost-effective analysis of unfixed cells . Calibrite beads allowed quantitation of live cells. Note: This staining protocol utilizes Zombie Aqua to assess live/dead status of cells prior to staining for surface markers, hence requiring cell suspension to be free of BSA in step 36 Alternatively, DAPI can also be used to identify live cells. with Zombie Aqua™ and analyzed without ˜xation (blue) or analyzed after ˜xation and permeabilization (red). They irreversibly conjugate to primary amine-containing proteins. 29. Zombie Violet dye has similar emission to BV421. Zombie Aqua™ Fixable Viability Kit (Biolegend) was used to label dead cells before staining, according to the manufacturer instructions. CD11c BV421 N418 BioLegend 117330 CD4 BV605 RM4-5 BioLegend 100547 Ly6c FITC HK1.4 BioLegend 128005 CD3e PerCp Cy5.5 145-2C11 BioLegend 100328 SIGLEC F PE E50-2440 BD Biosciences BD552126 Ly6G PE Cy7 1A8 BioLegend 127618 CD11b APC M1/70 BioLegend 101212 CD8a AF700 53-6.7 Thermo Fisher 56-0081-82 Interferon-regulatory factors (IRF)-1 Rabbit mAb clone: D5E4 and Phospho-NF-κB p65 clone Ser536 (Cell Signaling Technology, USA). Live Dead Marker Zombie Aqua, supplied by BioLegend, used in various techniques. For detection of cell surface NRP2 on human AMs, cells were incubated with goat anti-human NRP2 polyclonal goat IgG (AF-2215; R&D) or control goat Single-cell suspensions from either fresh human distal lung or primary organoid culture at approximately 4 weeks of culture were dissociated as above, treated with Fc Block (BioLegend), and incubated in FACS buffer with Zombie Aqua 1:400, 1 μg ml −1 PerCP-Cy5.5 anti-human EpCAM (CD326), 1 μg ml −1 APC anti-human ITGA6 (CD49f), 2 μg ml . zombie aqua fixable viability dye ( BioLegend ) 99 BioLegend zombie aqua fixable viability dye Zombie Aqua Fixable Viability Dye, supplied by BioLegend, used in various techniques. Following antibodies were used for the flow cytometry; LIVE/DEAD Zombie Aqua (BioLegend), anti-CD19 PE (HIB19, BioLegend), anti-CD3 PerCP/Cyanine 5.5 (SK7, BioLegend), anti-TCR gamma/delta Brilliant Violet 421 (B1, BioLegend), anti-CD45RA Brilliant Violet 785 (HI100, BioLegend), anti-CD56 FITC (HCD56, BioLegend), anti-CD19 Brilliant Violet 785 . The Zombie live/dead fixable dyes are amine-reactive fluorescent dyes that are impermeant to live cells but permeant to cells with compromised membranes. Subsequently, PBMCs were washed in 2% FCS in PBS and incubated for 30min on 4°C in 2% FCS in PBS supplemented with anti-CD56 Brilliant Violet 421 (Clone NCAM16.2; BD), anti-CD16 Product Details Bioz Stars score: 99/100, based on 1 PubMed citations. Zombie Aqua(TM) Fixable Viability Kit from BioLegend Supplier Page View Company Product Page Pricing Info .. Cells were cultured for 72 h in full medium in flat-bottom 96-well plates with stimulants as in cell survival assays, before harvesting and analysis on a flow cytometer. Cells were then permeabilized with 0.001% digitonin BioLegend provides DNA dyes, Propidium Iodide and 7- AAD, that enter and stain dead cells, but are impermeable to live cells for rapid, cost- effective analysis of unfixed cells. To reduce nonspecific binding, human cells were next blocked with normal mouse immunoglobulin G (IgG; Life Technologies; 60 μg/ml) in Flow Cytometry Staining Buffer . , according to the manufacturer instructions Fc ), Immunofluorescence Microscopy ( if ) Conjugate/Tag/Label NONAB_CHMPRB mounted, for. At 400 × g for 3 min at 4°C and discard the supernatant IFN. ( BDBiosciences ) withthecountperorgan or per ml of blood shown fixable dyes are amine-reactive fluorescent dyes that are impermeant live... 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