Cd34 apc clone 8g12

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The CD34 APC (clone 8G12) is a fluorescent-labeled antibody used to detect and identify CD34-positive cells in flow cytometry applications. CD34 is a transmembrane glycoprotein expressed on hematopoietic progenitor cells, and the antibody clone 8G12 is conjugated to the fluorochrome allophycocyanin (APC) for detection.

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CD34-APC (76 citations) CD34 (clone 8G12), (6 citations) CD34-APC (8G12) (2 citations) Anti-CD34 clone 8G12 APC (2 citations)

4 protocols using cd34 apc clone 8g12

Assessing Cell Proliferation and Apoptosis

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For the analysis of cell proliferation, Caco-2 and AML cells were stained with 5(6)-carboxyfluorescein diacetate N-succinimidyl ester (CFSE; Sigma-Aldrich) [26 (link)], CD34 APC (clone 8G12; BD Biosciences, New Jersey, USA) and CD45 PerCp (clone 2D1; BD Biosciences) before stimulation. Experiments were performed and analyzed in timer series experiments considering as paternal population for comparison the cells with highest CFSE intensity, according to landmark publications [27 (link),28 (link)].
For the analysis of apoptosis/necrosis, Caco-2 and AML cells were stained with FITC-annexin V (BD Biosciences) and propidium iodide (PI; Sigma-Aldrich). AML cells were also stained with CD34 APC (clone 8G12; BD Biosciences).
Proliferation and apoptosis analysis were performed after 4 days of co-culture with NETs in a FACScalibur flow cytometer (BD Biosciences). All data were analyzed with Flowjo V10.

Arelaki S., Arampatzioglou A., Kambas K., Papagoras C., Miltiades P., Angelidou I., Mitsios A., Kotsianidis I., Skendros P., Sivridis E., Maroulakou I., Giatromanolaki A, & Ritis K. (2016). Gradient Infiltration of Neutrophil Extracellular Traps in Colon Cancer and Evidence for Their Involvement in Tumour Growth. PLoS ONE, 11(5), e0154484.

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Immunophenotypic Surface Marker Analysis

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Immunophenotypic surface marker analysis was performed on a FACS Canto II (BD, Heidelberg, Germany) upon staining with CD14 allophycocyanin (APC, clone M5E2; BD), CD29 phycoerythrin (PE, clone MAR4; BD), CD31 PE (clone WM59; BD), CD34 APC (clone 8G12; BD), CD45 APC (clone HI30; BD), CD73 PE (clone AD2; BD), CD90 APC (clone 5E10; BD), CD105 fluorescein isothiocyanate (FITC, clone MEM-226; ImmunoTools, Friesoythe, Germany).

Fernandez-Rebollo E., Mentrup B., Ebert R., Franzen J., Abagnale G., Sieben T., Ostrowska A., Hoffmann P., Roux P.F., Rath B., Goodhardt M., Lemaitre J.M., Bischof O., Jakob F, & Wagner W. (2017). Human Platelet Lysate versus Fetal Calf Serum: These Supplements Do Not Select for Different Mesenchymal Stromal Cells. Scientific Reports, 7, 5132.

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Isolation and Characterization of Endothelial Cells

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Kidney perfusates were centrifuged at 800 RCF for 5 min. Later, all cells from kidney perfusate solution were washed in PBS and incubated for 10 min in darkness with FACS lysing solution (BD Biosciences, San Jose CA, USA) to remove red blood cells. The remaining cells were cultured in Endothelial Growth Media (EGM2; Lonza, Walkersville MD, USA). Culture media was refreshed every two days. Potential EC colonies were isolated by mechanically removing colonies with non-EC morphology under the microscope. After expansion, EC identity was confirmed by flow cytometry using the following anti-human antibodies: CD45-PerCP (clone 2D1; BD Biosciences), CD31-PECy7 (clone WM59; BioLegend, San Diego CA, USA), CD34-APC (clone 8G12, BD Biosciences), CD146-AmCyan (clone P1H12; BD Biosciences), CD133-BV421 (clone 7, BioLegend) and CD105-FITC (clone 266; BD Biosciences).

Tejeda-Mora H., Verhoeven J.G., Verschoor W., Boer K., Hesselink D.A., van den Hoogen M.W., van der Laan L.J., Baan C.C., Minnee R.C, & Hoogduijn M.J. (2021). Circulating endothelial cells transiently increase in peripheral blood after kidney transplantation. Scientific Reports, 11, 8915.

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Comprehensive Characterization of Mesenchymal Stem Cells

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Immunophenotypic surface marker analysis was performed on a FACS Canto II (BD, Heidelberg, Germany) upon staining with the following antibodies as described before: CD29-phycoerythrin (PE, clone MAR4; BD), CD34-APC (clone 8G12; BD), CD45-APC (clone HI30; BD), CD90-APC (clone 5E10; BD).
For adipogenic differentiation, MSC were cultured in medium supplemented with 1 mM dexamethasone, 10 mg/L insulin, 50 mM IBMX, and 0.2 mM indometacin. Twenty days after induction, cells were fixed with 40 g/L paraformaldehyde and lipid droplets were visualized by staining with Oil Red O solution [45 (link)]. Fluorescence microscopic pictures were taken from four randomly chosen areas with a Leica DM IL HC microscope (Leica, Wetzlar, Germany).
Osteogenic differentiation was induced with culture medium supplemented with 1 mM dexamethasone, 1 M β-sodium glycerophosphate, and 50 mM ascorbic acid and with medium changes every 3–4 days. After 40 days, osteogenic differentiation was analyzed by alkaline phosphatase staining. Alkaline phosphatase granules located in the cytoplasm stained intensively and the color was darker than the cell. Fluorescence microscopic pictures were taken from four randomly chosen areas with a Leica DM IL HC microscope (Leica Microsystems, Wetzlar, Germany).

Sun Y.P., Zhang B.L., Duan J.W., Wu H.H., Wang B.Q., Yu Z.P., Yang W.J., Shan Y.F., Zhou M.T, & Zhang Q.Y. (2014). Effect of NK4 Transduction in Bone Marrow-Derived Mesenchymal Stem Cells on Biological Characteristics of Pancreatic Cancer Cells. International Journal of Molecular Sciences, 15(3), 3729-3745.

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