For Cox8-mCherry-eGFP, Bio-Rad Ze5 Cell Analyzer was used. Cells were first sorted for mCherry positivity followed by eGFP. Analysis was done using FlowJo software. Flow cytometry analysis of immune cells was done using the Beckman Coulter MoFlo Astrios; immune cells from the colon were isolated by 25 mM EDTA digestion to remove epithelial cells, followed by a 0.5 mg/mL collagenase IV digestion, and were enriched for using a 40%–70% percoll gradient. Immune cells were stained for with CD45 APC eFluor 780, 1:200 (Invitrogen 47-0451-82); CD4 PECy7, 1:300 (eBioscience, 25-0041-82); CDllc FITC, 1:200 (BioLegend, 117305); CDllb APC, 1:250 (eBioscience, 17-0112-83); Ly6C V450, 1:300 (BD Biosciences, 560594), Ly6G PE, 1:300 (BD Biosciences, 560594), F4/80 BV510, 1:100 (BD Biosciences, 563633), 7AAD Percp Cy 5.5, 1:300 (BD Biosciences, 559925).
Ly6c v450
Manufactured by BD
The Ly6C V450 is a laboratory equipment product used for the detection and identification of specific cell surface antigens. It functions as a fluorescent-labeled antibody that binds to the Ly6C protein, allowing for the analysis of Ly6C-expressing cells through flow cytometry or other fluorescence-based methods.
Automatically generated - may contain errors
Lab products found in correlation
7aad percp cy 5, by BD (1 mentions)
Cd4 pe cy7, by Thermo Fisher Scientific (1 mentions)
Ly6g pe, by BD (1 mentions)
Moflo astrio, by Beckman Coulter (1 mentions)
Cd45 apc efluor780, by Thermo Fisher Scientific (1 mentions)
Ze5 cell analyzer, by Bio-Rad (1 mentions)
F4 80 bv510, by BD (1 mentions)
Cd45 v500, by BD (1 mentions)
Cd11b apc cy7, by BD (1 mentions)
F4 80 apc, by BD (1 mentions)
Cd3 pe cf594, by BD (1 mentions)
Ly6g pe cy7, by BD (1 mentions)
Fortessa flow cytometer, by BD (1 mentions)
Cxcr2 pe, by BD (1 mentions)
Efluor 780, by Thermo Fisher Scientific (1 mentions)
Cd11c pe cy7, by Thermo Fisher Scientific (1 mentions)
F4 80 pe, by BD (1 mentions)
Ly6g apc, by BD (1 mentions)
Anti mouse cd16 cd32 fc block, by BD (1 mentions)
Kaluza 1, by Beckman Coulter (1 mentions)
4 protocols using ly6c v450
1
Multicolor Flow Cytometry of Immune Cells
2
Aortic Inflammatory Cell Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Inflammatory cells in aortic vessels and blood were analyzed by flow cytometry as previously described.6 (link) Single-cell suspensions were treated with Fc block, washed, and stained with CD45 V500, CD3 PE-CF594, CD11b APC-Cy7, Ly6G PE-Cy7, Ly6C V450, F4/80 APC, and CXCR2 PE and their homologous isotype-matched negative controls (BD, Franklin Lakes, NJ). On the basis of a live gate, events were acquired on a Fortessa flow cytometer (BD) and analyzed.
3
Quantifying Immune Cell Populations in BAL
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell suspension of BAL was centrifuged at 4 °C, 2,900 rpm for 10 min, and cell viability was determined by incubation with fixable viability dye (FVD) eFluor780 (eBiosciences) for 20 min. For subsequent flow cytometry (Gallios; Beckman Coulter), cells were fixed with 4% paraformaldehyde (PFA) and then incubated with Fc block (anti-mouse CD16/CD32; BD Biosciences) on ice for 20 min, followed by surface marker staining with anti-mouse Abs: F4/80 (PE; BD), CD11b (PE-CF594; BD), CD11c (PE-Cy7; eBiosciences), Ly6G (APC; BD), and Ly6C (V450; BD). The data were analyzed using Kaluza 1.2 (Beckman Coulter). The absolute numbers of different cell types were calculated based on the proportion of viable events analyzed by flow cytometry as related to the total number of viable cells per sample.
4
Alveolar Macrophage Uptake of siRNA-Loaded Nanoparticles
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The ability of resident alveolar macrophages (rAM) to internalize NGs loaded with fluorescently-labeled siRNA (siCy5) was assessed by flow cytometry. Briefly, siCy5 was complexed with the lowest NG dose in previous experiment (100 µg) at a ratio of 1 pmol siCy5 per µg NGs. Mice were treated with free siRNA, siNGs, surfactant-coated siNGs or surfactantcoated siNGs-P (n=4). At various time points after treatment, BAL was collected as described above. First, high-affinity Fc receptors were blocked by incubation with purified anti-mouse CD16/CD32 (BD Biosciences) for 15 minutes at 4°C. Next, the rAM were selected based on their forward and side scatter pattern, intrinsic autofluorescence and their surface marker profile, using an antibody against CD11c (CD11c-PE-TR), SiglecF (SiglecF-PE) and Ly6C (Ly6C-v450; BD Biosciences). The samples were measured on a FACSLSRII TM flow cytometer (BD Biosciences). Data analysis was performed using the FlowJo TM analysis software (Treestar, Costa Mesa, USA).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!