The largest database of trusted experimental protocols

Percp conjugated cd19

Manufactured by BD
Sourced in United States

PerCP-conjugated CD19 is a fluorescent-labeled antibody that binds to the CD19 antigen, a cell surface marker expressed on B cells. It can be used in flow cytometry applications to identify and quantify B cell populations in biological samples.

Automatically generated - may contain errors

4 protocols using percp conjugated cd19

1

Breg Cell Enumeration by Flow Cytometry

Check if the same lab product or an alternative is used in the 5 most similar protocols
Evaluation of Breg cells frequencies by flow cytometry, circulating Breg cells were detected using FITC-conjugated-CD38, PE-conjugated-CD24 (BD Bioscience, USA), and PerCP-conjugated CD19 (BD Bioscience, USA).100 μl of blood sample was incubated with 10 μl of CD24, CD38, and CD19 for 20 minutes at 4 C in the dark. Following incubation, red blood cell lysis, washing, and analysis by FACS Calibur flow cytometry with Cell Quest software (Becton Dickinson Biosciences, USA) was done. An isotype-matched negative control was used for each sample. A Forward and side scatter histogram was used to define the lymphocytes. Then CD19+ B cells were gated. Then the expression of CD38 and CD24 on the CD19+B cells was revealed. Regulatory B cells were specified as CD19+CD24highCD38high cells (Figure 1).
+ Open protocol
+ Expand
2

Quantitative analysis of circulating Bregs

Check if the same lab product or an alternative is used in the 5 most similar protocols
Using flow cytometry, circulating Bregs were detected using FITC-conjugated-CD38, PE-conjugated-CD24 (Bioscience, USA), and PerCP-conjugated CD19 (BD Bioscience, USA). Briefly, 100 µl of blood sample was incubated with 10 µL of CD24, CD38 and CD19 for 20 min at 4 °C in the dark. Following incubation, RBCs were lysed and washed. Cells were fixed and permeabilized then stained with APC-conjugated IL-10 (BD Bioscience, San Jose, CA, USA) and analysis by FACS Calibur flow cytometry with CellQuest software (Becton Dickinson Biosciences, San Jose, CA, USA). An isotype-matched negative control was used for each sample. Forward and side scatter histograms were used to define the lymphocytes population. CD19+ IL-10+ B cells were gated, then the expression of CD38 and CD24 on the CD19+B cells were detected. Bregs were identified as CD19+ IL-10+CD24+hiCD38+hi cells. CD19+ B cells were selected based on the use of the isotype-matched negative control. However, for proper gating of the IL-10+ and CD24+hiCD38+hi cells, fluorescence minus one controls were employed, shown in light blue and red colors in the dot blots for the gated cells and controls, respectively (Figure 1A).
+ Open protocol
+ Expand
3

Characterization of Circulating B Lymphocytes and Bregs

Check if the same lab product or an alternative is used in the 5 most similar protocols
Circulating B lymphocytes and Bregs were detected in peripheral blood using the following markers (BD Biosciences, San Jose, CA, USA): PerCP-conjugated CD19, FITC-conjugated CD24 and PE-conjugated CD38. Briefly, the blood sample (100 μl) was incubated with 10 μL of the markers (CD19,CD24 and CD38) for 20 min at 4°C in the dark. RBCs were lysed using BD FACS Lysing solution (BD Biosciences, San Jose, CA, USA), and washing was done after incubation. Gating of CD19+ B cells was done, followed by CD38+ and CD24+ expression on CD19+ B cells. The analysis was done as described previously. CD19+CD24+hiCD38+hi cells were identified as Bregs. (Fig 3)
+ Open protocol
+ Expand
4

Flow Cytometric Identification of Regulatory B Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
Using flow cytometry, circulating Bregs were detected using PE-conjugated CD38, FITC-conjugated CD24 (Bioscience, USA) and PerCP-conjugated CD19 (BD Bioscience, USA).
One hundred microliters of the blood sample was incubated with 10 µL of CD24, CD38 and CD19 for 20 minutes at 4 C in the dark. Following incubation, red blood cells lysis, washing and analysis by FACS Calibur flow cytometry with CellQuest software (Becton Dickinson Biosciences, USA) were done. An isotype-matched negative control was used for each sample. Forward and side scatter histogram was used to define the lymphocytes population. Then, CD19+ B cells were gated. Then, the expression of CD38 and CD24 on the CD19+B cells was detected. Bregs were identified as CD19+CD24+hiCD38+hi cells (Figure 1).

Flow cytometric detection of regulatory B cells.

Notes: (A) Forward and side scatter histogram was used to define the lymphocytes population (R1). (B) The CD19+ cells (R2) were assessed within the lymphocyte population. (C) The expression of CD24 and CD38 was assessed in CD19+ cells to define CD19+CD24+highCD38+high cells (regulatory B cells).
+ Open protocol
+ Expand

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

Sign up now

Revolutionizing how scientists
search and build protocols!