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Cd34 bv650 clone 561

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The CD34-BV650 (clone 561) is a fluorochrome-conjugated antibody that binds to the CD34 antigen. CD34 is a cell surface glycoprotein expressed on hematopoietic progenitor cells and endothelial cells. This product can be used for the identification and enumeration of CD34-positive cells in flow cytometry applications.

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Lab products found in correlation

Cd11b pe cy7 clone icrf44, by BioLegend (2 mentions) Countbright beads, by Thermo Fisher Scientific (2 mentions) Cd15 af700 clone hi98, by BioLegend (2 mentions) Cd14 fitc clone m5e2, by BioLegend (2 mentions) Cd38 pe cy5 clone hit2, by BioLegend (2 mentions) Cytoflex lx, by Beckman Coulter (2 mentions) Cd3 apc clone hit3a, by BioLegend (1 mentions) Cd19 apc clone hib19, by BioLegend (1 mentions) Cd3 apc, by BioLegend (1 mentions) Pre designed guide rnas, by Integrated DNA Technologies (1 mentions) Guide rna, by Integrated DNA Technologies (1 mentions) Cas9 protein, by Integrated DNA Technologies (1 mentions) 4d nucleofector system, by Lonza (1 mentions)

3 protocols using cd34 bv650 clone 561

1

Myeloid Cell Enumeration from Plasma

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To assess the number of myeloid cells from plasma incubation experiments, cells were stained with the following panel: CD3-APC (clone HIT3a), CD19-APC (clone HIB19), CD56-APC (clone 5.1H11), CD14-FITC (clone M5E2), CD15-AF700 (clone HI98), CD11b-PE-Cy7 (clone ICRF44), CD34-BV650 (clone 561), and CD38-PE/Cy5 (clone HIT2) (BioLegend). After staining, cells were resuspended in FACS buffer with 2% CountBright beads (Invitrogen) to allow determination of absolute counts during analysis. Flow cytometry data were acquired on a Cytoflex LX (Beckman Coulter) and analyzed using FlowJo v10.1.
Reyes M., Filbin M.R., Bhattacharyya R.P., Sonny A., Mehta A., Billman K., Kays K.R., Pinilla-Vera M., Benson M.E., Cosimi L.A., Hung D.T., Levy B.D., Villani A.C., Sade-Feldman M., Baron R.M., Goldberg M.B., Blainey P.C, & Hacohen N. (2021). Plasma from patients with bacterial sepsis or severe COVID-19 induces suppressive myeloid cell production from hematopoietic progenitors in vitro. Science Translational Medicine, 13(598), eabe9599.
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2

Myeloid Cell Enumeration from Plasma

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To assess the number of myeloid cells from plasma incubation experiments, cells were stained with the following panel: CD3-APC (allophycocyanin) (clone HIT3a), CD19-APC (clone HIB19), CD56-APC (clone 5.1H11), CD14-FITC (clone M5E2), CD15-AF700 (clone HI98), CD11b-PE-Cy7 (clone ICRF44), CD34-BV650 (clone 561), and CD38-PE (phycoerythrin)/Cy5 (clone HIT2) (BioLegend). After staining, cells were resuspended in FACS buffer with 2% CountBright beads (Invitrogen) to allow determination of absolute counts during analysis. Flow cytometry data were acquired on a CytoFLEX LX (Beckman Coulter) and analyzed using FlowJo v10.1.
Reyes M., Filbin M.R., Bhattacharyya R.P., Sonny A., Mehta A., Billman K., Kays K.R., Pinilla-Vera M., Benson M.E., Cosimi L.A., Hung D.T., Levy B.D., Villani A.C., Sade-Feldman M., Baron R.M., Goldberg M.B., Blainey P.C, & Hacohen N. (2021). Plasma from patients with bacterial sepsis or severe COVID-19 induces suppressive myeloid cell production from hematopoietic progenitors in vitro. Science Translational Medicine, 13(598), eabe9599.
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3

CRISPR-Mediated Knockout of Cytokine Signaling Genes in CD34+ HSPCs

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Cas9 protein, predesigned guide RNAs targeting S100A8, S100A9, IL6ST, IL6R, IL10RA, and IL10RB, and nontargeting guide RNAs (from GeCKO v2 library) were purchased from Integrated DNA Technologies. Ribonucleoprotein complexes were assembled by combining 2.1 μl of 1X PBS, 1.2 μl of 100 μM guide RNA, and 1.7 μl of Cas9 protein (10 μg/ml) and incubating at room temperature for 15 min. The complexes were added to 50,000 to 100,000 CD34+ HSPCs resuspended in 20 μl of P3 (Lonza) and electroporated (program code DZ-100) using the 4D-Nucleofector system (Lonza). After electroporation, the cells were immediately transferred to 500 μl of HSPC media and rested for 48 hours. Knockout efficiency in CD34+ HSPCs was assessed after 48 hours via flow cytometry using the following panel: CD34-BV650 (clone 561), CD38-PE/Cy5 (clone HIT2), CD126-APC (clone UV4), and CD210-PECy7 (clone 3F9) (BioLegend).
Reyes M., Filbin M.R., Bhattacharyya R.P., Sonny A., Mehta A., Billman K., Kays K.R., Pinilla-Vera M., Benson M.E., Cosimi L.A., Hung D.T., Levy B.D., Villani A.C., Sade-Feldman M., Baron R.M., Goldberg M.B., Blainey P.C, & Hacohen N. (2021). Plasma from patients with bacterial sepsis or severe COVID-19 induces suppressive myeloid cell production from hematopoietic progenitors in vitro. Science Translational Medicine, 13(598), eabe9599.
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