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Mouse anti human cd19 apc

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Mouse anti-human CD19-APC is a fluorochrome-conjugated monoclonal antibody that binds to the CD19 cell surface antigen expressed on human B cells. It is a tool for the identification and enumeration of B cells in flow cytometric analysis.

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Lab products found in correlation

Goat anti human igg pe, by Southern Biotech (2 mentions) Mouse anti human cd81 pe, by BioLegend (2 mentions) Mouse anti human igm apc mhm 88, by Southern Biotech (2 mentions)

3 protocols using mouse anti human cd19 apc

1

Targeted Knockout Validation by Flow Cytometry

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Cell lines were transduced with sgRNA vectors marked by GFP. Three to four days after transduction, GFP levels were measured by flow cytometry on a BD FACS Calibur using CellQuest™ Pro version 6.0 and analyzed with FlowJo version 9. Cells were split every other day into doxycycline containing media and GFP levels were followed for 14 days and normalized to the day 0 measurement. All sgRNA and shRNA sequences are listed below. When surface proteins were targeted, knockout was validated by flow cytometry by spinning cells down, washing in FACS buffer (PBS plus 2% (vol/vol) FBS, 1mM EDTA), and stained at 4°C for 30 minutes in FACS buffer with fluorescently labeled antibodies: mouse anti-human CD19-APC (Biolegend SJ25C1, 1:500), mouse anti-human CD81-PE (Biolegend 5A6, 1:500), mouse anti-human IgM-APC (MHM-88), 1:400); or from Southern Biotech: goat anti-human IgG-PE (1:200).
Phelan J.D., Young R.M., Webster D.E., Roulland S., Wright G.W., Kasbekar M., Shaffer AL I.I.I., Ceribelli M., Wang J.Q., Schmitz R., Nakagawa M., Bachy E., Huang D.W., Ji Y., Chen L., Yang Y., Zhao H., Yu X., Xu W., Palisoc M.M., Valadez R.R., Davies-Hill T., Wilson W.H., Chan W.C., Jaffe E.S., Gascoyne R.D., Campo E., Rosenwald A., Ott G., Delabie J., Rimsza L.M., Rodriguez F.J., Estephan F., Holdhoff M., Kruhlak M.J., Hewitt S.M., Thomas C.J., Pittaluga S., Oellerich T, & Staudt L.M. (2018). A Multiprotein Supercomplex Controlling Oncogenic Signaling in Lymphoma. Nature, 560(7718), 387-391.
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2

Targeted Knockout Validation by Flow Cytometry

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Cell lines were transduced with sgRNA vectors marked by GFP. Three to four days after transduction, GFP levels were measured by flow cytometry on a BD FACS Calibur using CellQuest™ Pro version 6.0 and analyzed with FlowJo version 9. Cells were split every other day into doxycycline containing media and GFP levels were followed for 14 days and normalized to the day 0 measurement. All sgRNA and shRNA sequences are listed below. When surface proteins were targeted, knockout was validated by flow cytometry by spinning cells down, washing in FACS buffer (PBS plus 2% (vol/vol) FBS, 1mM EDTA), and stained at 4°C for 30 minutes in FACS buffer with fluorescently labeled antibodies: mouse anti-human CD19-APC (Biolegend SJ25C1, 1:500), mouse anti-human CD81-PE (Biolegend 5A6, 1:500), mouse anti-human IgM-APC (MHM-88), 1:400); or from Southern Biotech: goat anti-human IgG-PE (1:200).
Phelan J.D., Young R.M., Webster D.E., Roulland S., Wright G.W., Kasbekar M., Shaffer AL I.I.I., Ceribelli M., Wang J.Q., Schmitz R., Nakagawa M., Bachy E., Huang D.W., Ji Y., Chen L., Yang Y., Zhao H., Yu X., Xu W., Palisoc M.M., Valadez R.R., Davies-Hill T., Wilson W.H., Chan W.C., Jaffe E.S., Gascoyne R.D., Campo E., Rosenwald A., Ott G., Delabie J., Rimsza L.M., Rodriguez F.J., Estephan F., Holdhoff M., Kruhlak M.J., Hewitt S.M., Thomas C.J., Pittaluga S., Oellerich T, & Staudt L.M. (2018). A Multiprotein Supercomplex Controlling Oncogenic Signaling in Lymphoma. Nature, 560(7718), 387-391.
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3

Sorting Subsets of Human B Cells

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To sort the subsets of B cells, the isolated PBMCs were washed in PBS, blocked with 5% fetal bovine serum (FBS) for 30 min on ice, and then stained with mouse anti-human CD19 APC (BioLegend, 302211), mouse anti-human CD10 PerCP-eFluor (eBioscience, 46-0108-41), mouse anti-human CD27 FITC (eBioscience, 11-0279-41), mouse anti-human CD38 PE/Cy7 (eBioscience, 25-0388-41), and mouse anti-human IgM APC/Cy7 (BioLegend, 314519) for 30 min on ice in the dark. The stained cells were washed in PBS, and single cells were sorted by flow cytometry. Naive B cells were defined as CD38+/−CD10− cells gated on CD19+ cells. Memory B cells were defined as CD38++CD10+ cells gated on CD19+ cells. Plasma cells were defined as CD38+++CD10− cells gated on CD19+ cells. The B cells were sorted into 96-well plates by flow cytometric sorting to ensure that one cell was sorted into each well and avoid contamination. Each single B cell was carefully removed under a microscope with a mouth pipette and collected separately.
Shi Z., Zhang Q., Yan H., Yang Y., Wang P., Zhang Y., Deng Z., Yu M., Zhou W., Wang Q., Yang X., Mo X., Zhang C., Huang J., Dai H., Sun B., Zhao Y., Zhang L., Yang Y.G, & Qiu X. (2019). More than one antibody of individual B cells revealed by single-cell immune profiling. Cell Discovery, 5, 64.
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