Cd34 clone 8g12

Manufactured by BD

Sourced in United States

CD34 (clone 8G12) is a laboratory reagent used for the detection and identification of CD34-positive cells in biological samples. It is a monoclonal antibody that binds specifically to the CD34 antigen, which is expressed on the surface of hematopoietic stem and progenitor cells.

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Lab products found in correlation

Cd33 clone p67.6, by BD (2 mentions) Cd19 clone sj25c1, by BD (2 mentions) Cd45 clone 2d1, by BD (2 mentions) Facscanto 2, by BD (2 mentions) Cd45.2 clone 104, by Thermo Fisher Scientific (1 mentions) Ethylene diamine tetraacetic acid (edta), by Merck Group (1 mentions) Cd90 clone 5e10, by BD (1 mentions) Cd105 clone 266, by BD (1 mentions) Cd73 clone ad2, by BD (1 mentions) Cellquest pro software, by BD (1 mentions) Facsflow, by BD (1 mentions) Cd3 clone okt3, by BioLegend (1 mentions) Cd19 clone sj25c1, by BioLegend (1 mentions) Cd64 clone 10.1, by BioLegend (1 mentions) Cd10 clone hi10a, by BioLegend (1 mentions) Ficoll paque plus, by GE Healthcare (1 mentions) Facsaria 2, by BD (1 mentions) Cd45ra clone hi100, by Thermo Fisher Scientific (1 mentions) Fix perm kit, by Thermo Fisher Scientific (1 mentions) Facsaria fusion, by BD (1 mentions)

6 protocols using cd34 clone 8g12

Multiparametric Flow Cytometry and Histochemical Analysis of Murine AML

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For flow cytometry analyses, fluorescent antibodies against murine CD45.2 (clone 104, eBiosciences) and against human CD45 (clone HI30), CD33 (clone P67.6), CD34 (clone 8G12), CD19 (clone SJ25C1) (BD Biosciences), and CD45 (clone HI30, Invitrogen) were used. The analyses were performed on a FACS CantoII (BD Biosciences).
Humeri of fludarabine-treated SCID/beige mice transplanted with primary human AML blasts were fixed in 4% formaldehyde in phosphate buffer, decalcified in 10% EDTA (Sigma-Aldrich) and routinely processed for paraffin embedding. Serial 5 µm-thick sections were stained with Hematoxylin-Eosin and stained with anti-human CD33 (# NCL-L-CD33, clone PWS44, 1:100; Novocastra™) and Myeloperoxidase (#NCL-MYELO, clone 59A5, 1:100, Novocastra™) antisera.

Pievani A., Michelozzi I.M., Rambaldi B., Granata V., Corsi A., Dazzi F., Biondi A, & Serafini M. (2018). Fludarabine as a cost-effective adjuvant to enhance engraftment of human normal and malignant hematopoiesis in immunodeficient mice. Scientific Reports, 8, 9125.

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Immunophenotyping of MSCs after Au NP Exposure

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The immunophenotype of MSCs was analyzed after exposure to 10 nM Au NPs for 48 h. According to the recommendations of the International Society for Cellular Therapy [38 (link)] the following surface markers were measured: CD14 (clone M4P9, BD Biosciences, #345785), CD19 (clone SJ25C1, BD, #332780), CD34 (clone 8G12, BD, #345801), CD45 (clone 2D1, BD, #332784), CD73 (clone AD2, BD, #550257), CD90 (clone 5E10, BD, #559869), CD105 (clone 266, BD, #32830) and HLA-DR (clone B8.12.2, Immunotech, #PNIM0463U). In brief, MSCs were stained for 15 min at 4 °C with fluorochrome-labeled monoclonal antibodies, washed with PBS, and resuspended in FACSFlow™ (BD, #342003) with 3% formaldehyde (Merck, #103999). The samples were measured with a LSRII FCM device with CellQuest Pro™ Software (both BD). Isotype-matched antibodies were used as negative controls (BD, #342409, #347221, #345818). FCS data were analyzed with FlowJo™ software version 9.5.3 (TreeStar Inc).

Nold P., Hartmann R., Feliu N., Kantner K., Gamal M., Pelaz B., Hühn J., Sun X., Jungebluth P., del Pino P., Hackstein H., Macchiarini P., Parak W.J, & Brendel C. (2017). Optimizing conditions for labeling of mesenchymal stromal cells (MSCs) with gold nanoparticles: a prerequisite for in vivo tracking of MSCs. Journal of Nanobiotechnology, 15, 24.

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Isolation and Purification of CD34+ Hematopoietic Stem Cells

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For the CD34⁺ cells purification, bone marrow mononuclear cells were isolated by Ficoll-Paque Plus (GE HealthCare) density gradient centrifugation and stained using CD34 (clone 8G12; BD Biosciences) CD64 (clone 10.1; BioLegend) CD19 (clone SJ25C1; BioLegend) CD10 (clone HI10A; BioLegend) CD3 (clone OKT3; BioLegend) CD36 (clone CLB-IVC7; Sanquin Plesmanlaan) CD61 (clone RUU-PL7F12; BD Biosciences) for 15 min at RT. CD34⁺ CD64^- CD19^- CD10^- CD3^- CD36⁺ CD61⁺ cells were then sorted in a BD FACSAria II (BD Biosciences). Purified CD34⁺ cells were directly used for scRNA-seq analysis.

Ainciburu M., Ezponda T., Berastegui N., Alfonso-Pierola A., Vilas-Zornoza A., San Martin-Uriz P., Alignani D., Lamo-Espinosa J., San-Julian M., Jiménez-Solas T., Lopez F., Muntion S., Sanchez-Guijo F., Molero A., Montoro J., Serrano G., Diaz-Mazkiaran A., Lasaga M., Gomez-Cabrero D., Diez-Campelo M., Valcarcel D., Hernaez M., Romero J.P, & Prosper F. (2023). Uncovering perturbations in human hematopoiesis associated with healthy aging and myeloid malignancies at single-cell resolution. eLife, 12, e79363.

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Multiparameter Flow Cytometry for Studying HIV-1

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Antibodies to the following human proteins were used for flow cytometry: CD4 (clone RPA-T4, BrilliantBlue 515-conjugated, BD Bioscience, Franklin Lakes, NJ, USA), CD34 (clone 8G12, allophycocyanin (APC)-conjugated, BD Bioscience), CD389 (clone HB7, V450-conjugated, BD Bioscience), CD45RA (clone HI100, phycoerythrin (PE)-Cyanin7 (Cy7)-conjugated, Thermo Fisher Scientific), CD90 (Thy-1) (clone eBio5E10, PerCP-Cy5.5-conjugated, Thermo Fisher Scientific), CD184 (CXCR4, clone 12G5, PE-conjugated, Miltenyi Biotec), CD195 (CCR5, clone REA245, PE-conjugated, Miltenyi Biotec), CD3 (clone BW264/56, APC-conjugated, Miltenyi Biotec), and HIV-1 core antigen (p24) (clone KC-57, RD1-conjugated, Beckman Coulter). The secondary reagent used was streptavidin (APC-eFlour^®780-conjugated, Thermo Fisher Scientific) to fluorescently label biotinylated lineage antibodies in the Lin-depletion kit in order to exclude remaining Lin⁺ cells via FACS. For HIV-1 p24 antigen staining, cells were fixed and permeabilized using the FIX & PERM^® kit according to the manufacturers’ instructions (Thermo Fisher Scientific). Samples were analyzed using a BD FACSCanto II after fixation (3% formaldehyde). Cell sorting was performed using a BD FACSAria Fusion (all BD Bioscience) flow cytometer. Compensation was performed using single-stained Lin⁺ cells for each marker.

Renelt S., Schult-Dietrich P., Baldauf H.M., Stein S., Kann G., Bickel M., Kielland-Kaisen U., Bonig H., Marschalek R., Rieger M.A., Dietrich U, & Duerr R. (2022). HIV-1 Infection of Long-Lived Hematopoietic Precursors In Vitro and In Vivo. Cells, 11(19), 2968.

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Multicolor Flow Cytometry Immunophenotyping

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After thawing, cells were stained with directly labeled antibodies recognizing CD33 (clone P67.6; PE-conjugated), CD3 (clone SK7; PerCP-conjugated), CD45 (clone 2D1; FITC-conjugated), PD-L1 (clone MIH1, PE-conjugated), PD-L2 (clone MIH18, BV650-conjugated), CD86 (clone 2331, APC-conjugated), CD34 (clone 8G12; APC-conjugated; all from BD Biosciences) and CD80 (clone 2D10, PE-Cy7-conjugated; from BioLegend). To identify nonviable cells, samples were stained with DAPI. At least 10 000 events were acquired on an LSRII flow cytometer (BD Biosciences), and DAPI^- cells were analyzed using FlowJo.

Laszlo G.S., Gudgeon C.J., Harrington K.H, & Walter R.B. (2015). T-cell ligands modulate the cytolytic activity of the CD33/CD3 BiTE antibody construct, AMG 330. Blood Cancer Journal, 5(8), e340-.

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Multiparameter Flow Cytometry Assay

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Cells were stained with directly labeled antibodies recognizing CD33 (clone P67.6), CD3 (clone SK7), CD34 (clone 8G12), and CD38 (clone HB7; all from BD Biosciences, San Jose, CA) and CD45 (clone HI30; eBioscience). The activities of P-glycoprotein (Pgp), multidrug resistance protein (MRP), and breast cancer resistance protein (BCRP) were determined using a commercial flow cytometric kit (eFLUXX-ID, ENZO Life Sciences, Plymouth Meeting, PA). To identify nonviable cells, samples were stained with 4',6-diamidino-2-phenylindole (DAPI). At least 10,000 events were acquired on a Canto II flow cytometer (BD Biosciences), and DAPI^- cells analyzed using FlowJo (Tree Star, Ashland, OR).

Harrington K.H., Gudgeon C.J., Laszlo G.S., Newhall K.J., Sinclair A.M., Frankel S.R., Kischel R., Chen G, & Walter R.B. (2015). The Broad Anti-AML Activity of the CD33/CD3 BiTE Antibody Construct, AMG 330, Is Impacted by Disease Stage and Risk. PLoS ONE, 10(8), e0135945.

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