Mice (WT and Gsdmd-/-) were intravenously challenged with 3 × 105 CFU WT (GFP+) or candidalysin-deficient (ece1Δ/Δ) C. albicans. Three days after C. albicans infection, kidneys were homogenized using 40 µm cell strainers. Red blood cells were lysed with 1 ml ACK lysis buffer (Gibco) for 5 min at room temperature. Cells were then stained with PE-Cy7-conjugated CD45 (BioLegend), PE-conjugated F4/80 (BioLegend), and APC-conjugated CD11b (BioLegend) antibodies for 30 min. For cells isolated from WT (GFP+) C. albicans-infected mice, macrophages containing trapped C. albicans were recognized by GFP fluorescence. For cells isolated from candidalysin-deficient (ece1Δ/Δ) C. albicans-infected mice, the samples were fixed and permeabilized (BD Biosciences) after surface marker staining, and then stained with FITC-conjugated anti-C. albicans antibody (thermofisher) for 1 h. Flow cytometry was performed on the LSRFortessa (BD Biosciences) instrument. Flow cytometry data were analyzed with FlowJo software.
Pe cy7 conjugated cd45
Manufactured by BioLegend
Sourced in United States
The PE-Cy7-conjugated CD45 is a flow cytometry reagent used to detect the CD45 antigen expressed on the surface of various hematopoietic cell types. It is a fluorescent-labeled monoclonal antibody that can be used to identify and analyze these cells in a sample.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using pe cy7 conjugated cd45
1
Quantifying C. albicans Infection in Mice
2
Isolation and Characterization of GFP-Labeled Mesenchymal Stem Cells from Synovium
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
1 Â 10 6 GFP þ MSCs were injected 1week after the ACLT, then synovium was harvested 1day after the injection, following digestion with collagenase V for three hours. After filtering through a 70 mm cell strainer and centrifuging at 1500 rpm for 5 min, cells were suspended in ice-cold Hank's balances salt solution and then stained for 30 min on ice with a monoclonal antibody of APCconjugated CD90 (Becton, Dickinson and Company, Franklin Lakes, NJ, USA). Synovial fluid was also extracted from the same knee, and prepared in the same manner without collagenase digestion. Propidium iodide (PI) fluorescence was measured, and a live cell gate was defined that excluded the cells positive for PI. Additional gates were defined as positive for GFP and CD90. Double positive cells were further analyzed for PE-conjugated CD29, PEconjugated CD31, and PEcy7-conjugated CD45 (Biolegend, San Diego, CA, USA). Flow-cytometric analysis and sorting were performed on a MoFlo (Beckman Coulter, Brea, CA, USA), and the data were analyzed using FlowJo software (Tree Star, Ashland, OR, USA) 26 .
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!