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Cd34 pe

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CD34-PE is a fluorochrome-conjugated monoclonal antibody that binds to the CD34 antigen. CD34 is a transmembrane glycoprotein expressed on hematopoietic stem and progenitor cells. The PE fluorochrome allows for the detection and analysis of CD34-positive cells using flow cytometry.

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Lab products found in correlation

Cd90 pe, by BioLegend (13 mentions) Cd45 pe, by BioLegend (10 mentions) Cd105 pe, by BioLegend (7 mentions) Cd73 pe, by BioLegend (6 mentions) Cd45 fitc, by BioLegend (5 mentions) Cd19 pe, by BioLegend (5 mentions) Cd90 fitc, by BioLegend (5 mentions) Hla dr pe, by BioLegend (4 mentions) Cd105 fitc, by BioLegend (4 mentions) Cd29 pe, by BioLegend (4 mentions) Flow cytometry, by BD (4 mentions) Cd14 pe, by BioLegend (4 mentions) Cd45 apc, by BioLegend (4 mentions) Cd73 fitc, by BioLegend (3 mentions) Cd44 fitc, by BioLegend (3 mentions) Fetal bovine serum (fbs), by Atlanta Biologicals (3 mentions) Qbsf 60, by Quality Biological (3 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (3 mentions) Cd10 pacific blue, by BioLegend (3 mentions) Cd19 pe cy5, by BioLegend (3 mentions)

Spelling variants of this product

Anti-CD34-PE (26 citations) PE anti-mouse CD34 (5 citations) Anti-CD34 PE-Cy7 (4 citations) Anti-human CD34-PE (4 citations) PE anti-human CD34 Antibody (3 citations) Anti human CD34 PE-Cy7 (3 citations) Anti-CD34 PE antibody (2 citations) PE‐anti mouse CD34 antibody (2 citations) CD34-PE antibody (2 citations) PE conjugated anti-CD34 and anti-CD45 primary antibodies (2 citations)

42 protocols using cd34 pe

1

ECFC and MSC Immunophenotyping Protocol

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ECFC single-cell suspension was generated by detaching cells with TrypLE™ Express Enzyme (Gibco, USA) and resuspended to a concentration of 1 × 107 cells/ml. Samples were incubated, respectively, with anti-human CD31-FITC (eBioscience, USA), VEGFR2/KDR-PE (R&D, USA), CD144-FITC (Abcam, UK), CD34-PE (Biolegend, USA), CD45-FITC (Biolegend, USA), and CD14-FITC (eBioscience, USA). MSCs were resuspended to a concentration of 1 × 106 cells/ml and incubated, respectively, with anti-human CD29-PE (Biolegend, USA), CD90-PE (Biolegend, USA), CD14-FITC (Biolegend, USA), CD19-PE (Biolegend, USA), CD73-FITC (Biolegend, USA), CD105-FITC (Biolegend, USA), HLA-DR-PE (Biolegend, USA), CD34-PE (Biolegend, USA), CD45-FITC (Biolegend, USA), and CD31-FITC (eBiosciences, USA). 5 μl antibody solution was added into 100 μl cell suspension and incubated for 30 minutes at 4°C in the dark; 400 μl of PBS was added and cells were analyzed with FACSAria I (Becton Dickinson, USA) or Accuri C6 (Becton Dickinson, USA) and Becton Dickinson CELLQuest software.
Lu H., Wang F., Mei H., Wang S, & Cheng L. (2018). Human Adipose Mesenchymal Stem Cells Show More Efficient Angiogenesis Promotion on Endothelial Colony-Forming Cells than Umbilical Cord and Endometrium. Stem Cells International, 2018, 7537589.
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2

Flow Cytometric Analysis of DPSC Markers

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Flow cytometry was performed to detect human DPSCs surface markers. Young and aging DPSCs were collected, washed with phosphate-buffered saline (PBS), and fixed in 4% paraformaldehyde (PFA). After fixation for 30 min, the cells were permeabilized and incubated with antibodies for 1 h at 4°C. The following antibodies specific for human surface antigens were used: CD44-FITC (Biolegend, 103005), CD90-FITC (Biolegend, 328107), CD105-PE (eBioscience, 4300023), CD11b-FITC (eBioscience, 4271325), CD14-FITC (Biolegend, 301803), CD34-PE (Biolegend, 119307), and CD45-FITC (Biolegend, 304005). The corresponding isotype-matched (IgG) antibodies conjugated to PE and FITC served as negative controls. Data were analyzed using the FlowJo software (FlowJo, Ashland, OR, USA).
Zhang S., Zhang R., Qiao P., Ma X., Lu R., Wang F., Li C., E L, & Liu H. (2021). Metformin-Induced MicroRNA-34a-3p Downregulation Alleviates Senescence in Human Dental Pulp Stem Cells by Targeting CAB39 through the AMPK/mTOR Signaling Pathway. Stem Cells International, 2021, 6616240.
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3

Reactive Oxygen Species Detection Assay

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Reactive Oxygen Species Detection Assay Kit was purchased from BioVision (San Francisco, CA, USA). Wogonin was purchased from MedChemExpress (Israel Shekel), and LPS was purchased from Sigma-Aldrich (St. Louis, MO, USA). The following antibodies were used for cell surface staining: CD34-PE, CD44-FITC, CD29-PE-Cy7, Sca-1-AF700, CD45-AF700, and CD49e-APC are all from BioLegend and Mouse IL-10 ELISA Set and Zombie Green™ Fixable Viability Kit were purchased from BD Biosciences. Mouse IL-10 MAb was purchased from R&D Systems.
Wu Q., Xie S., Zhu Y., Chen J., Tian J., Xiong S., Wu C., Ye Y, & Peng Y. (2021). Wogonin Strengthens the Therapeutic Effects of Mesenchymal Stem Cells in DSS-Induced Colitis via Promoting IL-10 Production. Oxidative Medicine and Cellular Longevity, 2021, 5527935.
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4

CD34+ Hematopoietic Progenitor Differentiation

Cited 4 times
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Unstimulated and TLR-Stimulated CD34+ hematopoietic progenitors were plated at 10,000 cells/well for liquid, stromal cell-free B lineage cultures, or 1000 cells per sample in Methocult H4434 Classic (Stemcell Technologies), according to the manufacturer’s directions, as previously described [4 (link),5 (link),18 (link),19 (link)]. For B lineage liquid culture, cells were grown in QBSF®60 (Quality Biological, Inc.) supplemented with 10% FBS (Atlanta Biologicals), 100U Penicillin/Streptomycin (Gibco), 10% hASC conditioned media (LaCell LLC), 10 ng/ml stem cell factor, 10 ng/ml granulocyte stimulating factor, 5 ng/ml FLT-3 ligand, and 5 ng/ml IL-7 (as above). Half of the media was replaced once a week. After 4 weeks, cells were harvested, counted and assessed by flow cytometry using CD34-PE, CD10-Pacific Blue, CD19-PE-Cy5 (BioLegend), CD33-APC (BD Biosciences), and goat anti-ARID3a with a FITC-conjugated anti-goat secondary. Methocult cultures were incubated for 14 days and colonies were analyzed visually for monocytic, granulocytic and erythrocytic lineage cell colonies and counted using a Nikon Eclipse TS100 inverted microscope. Methocult cultures were harvested with warm 1X PBS, washed twice with warm PBS and once with ice cold PBS and then assessed by flow cytometry for the following surface markers: CD34, CD33, CD14, CD16, and CD41 with intracellular ARID3a.
Ratliff M.L., Shankar M., Guthridge J.M., James J.A, & Webb C.F. (2020). TLR engagement induces ARID3a in human blood hematopoietic progenitors and modulates IFNα production. Cellular immunology, 357, 104201.
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5

Stem Cell Immunophenotyping by Flow Cytometry

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The 3rd generation of stem cells was harvested, and washed once with phosphate-buffered saline(PBS). Cells (1 × 105 per sample) were stained with the following mouse anti-human monoclonal antibody at room temperature for 30 min: anti-CD44-FITC, -CD105-FITC, -CD29-PE, -CD34-PE (Biolegend, San Diego, CA, USA), isotype-identical antibody (Biolegend) served as controls. The cells were washed with PBS two times, and then resuspended in PBS. Fluorescent labeling was analyzed by flow cytometric analysis and CellQuest Pro software (BD Bioscience, Franklin Lakes, NJ, USA).
Wang Y., Wang F., Zhao H., Zhang X., Chen H, & Zhang K. (2014). Human Adipose-Derived Mesenchymal Stem Cells Are Resistant to HBV Infection during Differentiation into Hepatocytes in Vitro. International Journal of Molecular Sciences, 15(4), 6096-6110.
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6

Isolating Inner Ear Cell Populations

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Math1-GFP mice and CD-1 mice were euthanized at P1 and P2, respectively, and their temporal bone removed. Inner ear tissues were dissected and placed into 0.5 mg/ml Thermolysin (Sigma-Aldrich Cat# T7902) for 20 minutes at 37°C – 5% CO2. The Thermolysin was then replaced with Accutase (Sigma-Aldrich Cat# A6964) followed by three rounds of 3 minutes at 37°C and mechanical dissociation. The Accutase was inactivated with IMDM (Sigma-Aldrich, Cat# I6529) supplemented with 5% fetal bovine serum and the cell suspension was filtered through a 35-μm cell strainer to eliminate cell clumps. Dissociated cells were stained with CD326- APC (1:2,000; BioLegend Cat# 118213, RRID:AB_1134105), CD49falexa488 (1:100; BioLegend Cat# 313607, RRID:AB_493634), and CD34-PE (1:200; BioLegend Cat# 128609, RRID:AB_2074602) before FACS.
Hertzano R., Gwilliam K., Rose K., Milon B, & Matern M.S. (2020). Cell Type–Specific Expression Analysis of the Inner Ear: A Technical Report. The Laryngoscope, 131(Suppl 5), S1-S16.
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7

Multicolor Flow Cytometry Immunophenotyping

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Cultured cells were stained in PBS supplemented with 2% fetal bovine serum (FBS) at 4 °C for 30 min with the following human antibodies: CD34 PE (Biolegend, clone 581, Santiago, CA, US), CD34 FITC (BD Biosciences, clone 581, Franklin Lake, NJ, US), CD38 PE-Cy7 (BD Biosciences, clone HIT2), CD90 APC (BD Biosciences, clone 5E10), CD45 PE (Biolegend, clone HI30), CD11b/Mac1 APC (BD Biosciences, clone ICRF44), CD3 FITC (BD Biosciences, clone HIT3a), CD19 BV201 (Biolegend, clone SJ25C1), anti-mouse CD45 PErCP-Cy5.5 (Biolegend, clone 30-F11) and 7-amino-actinomycin D (7-AAD) (Biolegend). 7-AAD was used to exclude dead cells. Stained cells were washed once with PBS supplemented with 2% FBS and analyzed using the BD LSRFortessa (BD Biosciences). Proportion of positive/negative cells with the same mean fluorescence intensity (MFI) was represented.
Jiang M., Chen H., Lai S., Wang R., Qiu Y., Ye F., Fei L., Sun H., Xu Y., Jiang X., Zhou Z., Zhang T., Li Y., Xie J., Fang Q., Gale R.P., Han X., Huang H, & Guo G. (2018). Maintenance of human haematopoietic stem and progenitor cells in vitro using a chemical cocktail. Cell Discovery, 4, 59.
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8

Quantification of CD34+mono Cells in Peripheral Blood

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Peripheral blood samples were obtained from donors at a median of 4 days after G-CSF administration (range: 3–6). Mononuclear cells were separated by density gradient sedimentation using Lymphoprep™ (Axis-Shield PoC AS), and cryopreserved at −80 °C until use. Cells were incubated with anti-human Lin (Lineage: CD3, CD19, CD56)-FITC, CD34-PE, CD33-APC or CD83-APC, CD11b-PECy7, CD14-APCCy7 monoclonal antibodies, and 7AAD (BioLegend). Data were analysed with FACSverse and FACSuite (BD Biosciences).
CD34+mono was defined as Lin−CD34highCD14+CD11b+CD33+ cells (Fig. 1). When ≥0.1% of CD34+mono among CD34-positve cells was detected with ≥5 events, the donor was considered to be positive for CD34+mono.
Nakasone H., Kikuchi M., Kawamura K., Akahoshi Y., Sato M., Kawamura S., Yoshino N., Takeshita J., Yoshimura K., Misaki Y., Gomyo A., Tanihara A., Kusuda M., Tamaki M., Kimura S.I., Kako S, & Kanda Y. (2019). Increased CD83 expression of CD34-positive monocytes in donors during peripheral blood stem cell mobilization in humans. Scientific Reports, 9, 16499.
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9

Adipose-Derived Stem Cell Isolation and Characterization

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Similar to the aforementioned fibroblast extraction process, after removing vascular tissues, adipose tissues were minced and digested with 4 mg/mL type I collagenase for 20 min. Subsequently, the mixture was allowed to sit for 3–5 min before removing the supernatant. The supernatant was centrifuged to obtain primary ADSC, and proliferation was continued in the culture with Medium Essential Medium alpha (MEM-α; BI) containing 10% FBS. To identify surface markers of ADSC, the fluorescence-conjugated antibodies CD105-PE, CD73-PE, CD90-PE, CD34-PE, and CD45-PE (BioLegend, San Diego, CA, USA) were incubated with three sub-passages of ADSC several times and analyzed using flow cytometer (BD Pharmingen, San Diego, CA, USA). To identify the differentiation potential, ADSCs were cultured for several days, according to the instructions of the adipogenic (Cyagen, Santa Clara, CA, USA) and osteogenic (Cyagen) induction kits. We then visualized the differentiation results after Oil Red O (Solarbio) or Alizarin Red S (Solarbio) ing according to corresponding instructions under a microscope. We then used an alkaline phosphatase (ALP) kit (BI) to detect the stem cell properties of ADSCs. Finally, a light microscopic (Ningbo ShunYu Instruments Co., Ltd., Ningbo, China) analysis was performed.
Shao Z., Xu J., Xu X., Wang X., Zhou Y., Li Y, & Li K. (2023). Exosomes Derived from Human Adipose Mesenchymal Stem Cells Inhibits Fibrosis and Treats Oral Submucous Fibrosis via the miR-181a-5p/Smad2 Axis. Tissue Engineering and Regenerative Medicine, 21(1), 123-135.
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10

Characterization of BM-MSCs Prior to Infusion

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BM‐MSCs were characterized just before infusion, by cell surface expression of CD73‐PE, CD90‐PE, CD19‐PE, CD34‐PE, and CD45‐PE (BioLegend, San Diego, CA); CD105‐PE, HLADR‐PE, and CD14‐PE (Becton Dickinson); propidium iodide (PI; Sigma); and isotype IgG1‐PE and isotype IgG2a‐PE (Becton Dickinson). For immunolabeling, cells were incubated for 45 minutes at 2°C–8°C. PI‐negative BM‐MSCs (viable) were characterized using FC500 flow cytometer (Beckman Coulter, Mississauga, ON, Canada) and analyzed using FlowJo (Treestar).
Chahal J., Gómez‐Aristizábal A., Shestopaloff K., Bhatt S., Chaboureau A., Fazio A., Chisholm J., Weston A., Chiovitti J., Keating A., Kapoor M., Ogilvie‐Harris D.J., Syed K.A., Gandhi R., Mahomed N.N., Marshall K.W., Sussman M.S., Naraghi A.M, & Viswanathan S. (2019). Bone Marrow Mesenchymal Stromal Cell Treatment in Patients with Osteoarthritis Results in Overall Improvement in Pain and Symptoms and Reduces Synovial Inflammation. Stem Cells Translational Medicine, 8(8), 746-757.
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