The immune infiltrate present into the bladder was analyzed by flow cytometry as previously described.5 (link) Briefly, bladders were minced using a scalpel followed by digestion with RPMI medium supplemented with 5% FBS containing 0.5 mg/mL collagenase II (Sigma, Spain) and 1 U/mL DNAse I at 37°C for two-three successive 30 min cycles, with continuous shaking. The cell suspension obtained was filtered through a 40-μm disposable plastic strainer (Becton & Dickinson) and pelleted for staining. The cells were labeled with the following antibodies: PerCP-CD45 APC-Cy7-CD3, FITC-CD4, Alexa 700-CD8, APC-CD62L and BV786-CD127, PE-Dazzle-CD44 and FITC-CD45R/B220 (Biolegend). The lymphocyte gate was defined by morphological parameters, and dead cells were excluded using a live/dead fixable Aqua Dead Cell Stain kit (Invitrogen). Samples were acquired in a Fortessa flow cytometer (Becton & Dickinson), and the data were analyzed using FlowJo software (9.8 v; TreeStar, Portland, OR, USA). Absolute cell numbers were obtained by using Perfect-Count Microspheres (Cytognos). Gating strategy is shown in Supplementary Figure 3.
Fitc cd45r b220
Manufactured by BioLegend
FITC-CD45R/B220 is a fluorescently-labeled antibody that specifically binds to the CD45R/B220 antigen expressed on the surface of B cells and certain subsets of T cells. It can be used for the identification and characterization of these cell populations in various applications, such as flow cytometry.
Automatically generated - may contain errors
Lab products found in correlation
Cd4 fitc, by BioLegend (2 mentions)
Fortessa flow cytometer, by BD (1 mentions)
Aqua dead cell stain kit, by Thermo Fisher Scientific (1 mentions)
Cd62l apc, by BioLegend (1 mentions)
40 μm disposable plastic strainer, by BD (1 mentions)
Alexa 700 cd8, by BioLegend (1 mentions)
Collagenase 2, by Merck Group (1 mentions)
Dnase 1, by Merck Group (1 mentions)
Perfect count microspheres, by Cytognos (1 mentions)
Cd279 pd 1 pe, by BioLegend (1 mentions)
2 protocols using fitc cd45r b220
1
Flow Cytometry Analysis of Bladder Immune Infiltrate
2
RABV Strain LBNSE Characterization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Female ICR mice were purchased from the laboratory animal center of Huazhong Agricultural University, Wuhan, China. The recombinant RABV strain LBNSE was derived from the Street Alabama Dufferin (SAD)‐L16, which is widely applied for the development of vaccine.18 LBNSE with two mutations in the G protein, N194K and R333E, was proliferated in BSR cells.18 , 19 The rabies challenge virus strain‐11 (CVS‐11) was proliferated in BSR cells, too. The BSR cells, a cloned cell line come from BHK‐21 cells, were cultured at 37°C in Dulbecco's modified Eagle's medium (DMEM) (Mediatech, USA) containing 10% fetal bovine serum (FBS, Gibco) and antibiotics (penicillin‐streptomycin solution, 100×) (Beyotime, Wuhan, China). Fluorescein isothiocyanate (FITC)‐conjugated the antibodies against the RABV‐nucleoprotein (N) were obtained from FujiRab (Melvin, PA). Samples of dLNs were stained with monoclonal antibodies for flow cytometry, including FITC‐CD4 (BioLegend), APC‐CD185 (CXCR5) (BioLegend), PE‐CD279 (PD‐1) (BioLegend), FITC‐CD45R/B220 (BioLegend), Alexa Fluor 647‐GL7 (BioLegend), and PE‐CD95 (APO‐1/Fas) (eBioscience).
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!