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Fitc anti human cd34

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FITC anti-human CD34 is a fluorescently-labeled antibody that binds to the CD34 protein expressed on the surface of hematopoietic stem and progenitor cells. It is used in flow cytometry applications to identify and quantify CD34-positive cells.

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Lab products found in correlation

Facscalibur, by BD (4 mentions) Staining buffer, by BD (2 mentions) Cellquest software, by BD (2 mentions) Apc anti human cd206, by BioLegend (2 mentions) Alexa fluor 488 anti human cd105, by BioLegend (2 mentions) Pe anti human cd45, by BioLegend (2 mentions) Pe anti human cd206, by BioLegend (2 mentions) Pe anti human cd86, by BioLegend (2 mentions) Apc anti human cd86, by BioLegend (2 mentions) Accutase, by Nacalai Tesque (2 mentions) Human trustain fcx fc receptor blocking solution, by BioLegend (2 mentions) Alexa fluor 488 anti human cd11b, by BioLegend (2 mentions) Fitc anti human cd90, by BioLegend (2 mentions) Fitc anti human cd73, by BioLegend (2 mentions) Anti mouse cd45 fitc, by Thermo Fisher Scientific (1 mentions) Anti human cd45 apc, by Thermo Fisher Scientific (1 mentions) Annexin 5 fitc apoptosis detection kit, by 4A Biotech (1 mentions) Fibronectin, by BD (1 mentions) Mouse igg2a fitc, by BioLegend (1 mentions) Mouse igg1k pe, by BioLegend (1 mentions)

Close spelling variants of this product

CD34-FITC (32 citations) Anti-CD34 (26 citations) Anti-CD34-FITC (10 citations) FITC-conjugated anti-human CD34 (2 citations) Fluorescein isothiocyanate (FITC) anti-human CD34 antibodies (2 citations)

10 protocols using fitc anti human cd34

1

Comprehensive Cell Analysis Protocol

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FACS analysis was performed routinely by using a CaliburTM flow cytometer (BD Immunocytometry Systems). Anti-mouse CD45-FITC (#104, eBioscience), anti-human CD45-APC (HI30, eBioscience), anti-human CD34-FITC (#581, Biolegend). Cell-cycle analysis was performed using DNA binding dye propidiumiodide (PI). Hematopoietic cells were fixed in 50 % ethanol and resuspended to 0.2 mL of 10 mg/mL RNAaseA and 50 µg/mL PI. Cell-cycle kinetics was performed with routine protocols using the FACS Calibur flow cytometer (Becton–Dickinson, CA). Apoptosis was analyzed by using an Annexin V-FITC Apoptosis Detection Kit (4A Biotech, Beijing, China).
Tian D.M., Liang Y.M, & Zhang Y.Q. (2016). Endothelium-targeted human Delta-like 1 enhances the regeneration and homing of human cord blood stem and progenitor cells. Journal of Translational Medicine, 14, 5.
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2

Endothelial Cell Migration Assay

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Vacutainers for blood collection -K2-EDTA (Cat: 367863), Vacutainers for serum separation (Cat: 367812), fibronectin (Cat: 356008) and matrigel (Cat: 356231) were purchased from Becton Dickinson Biosciences. EGM-2 Bullet kits (Cat: CC-4176) were purchased from Lonza. Fluorescent antibodies, anti-human CD34-FITC (Cat: 343603) and anti-human VEGFR3-PE (Cat: 356203), and isotype control antibodies, mouse IgG2a-FITC (Cat: 400207) and mouse IgG1k-PE (Cat: 400113), were purchased from BioLegend. Transwell thin inserts (Cat: 3422) with 8.0 lm pore size were from Costar Corning. Multi-Analyte ELISA Array Kit was from Qiagen (Cat: MEH-004A). Trizol from Invitrogen (Cat: 15596-018), human SDF-1a (Cat: S190), gelatin (Cat: G1393), collagen (Cat: C0543), histopaque (Cat: 10771) and other dry chemicals were from Sigma-Aldrich. All qRT-PCR primers were purchased from Integrated DNA Technologies (IDT).
Nathan A.A., Dixit M., Babu S, & Balakrishnan A.S. (2017). Comparison and functional characterisation of peripheral blood mononuclear cells isolated from filarial lymphoedema and endemic normals of a South Indian population. Tropical medicine & international health : TM & IH, 22(11).
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3

Immunophenotyping of GMSCs and Macrophages

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The expression of CD73, CD90, CD105, CD34, CD45, and CD11b in GMSCs and CD11b, CD206, and CD86 in PBMC-derived macrophages were analyzed using the FACS Calibur (Becton Dickinson, USA) and the CellQuest software (Becton Dickinson). The adherent cells were washed with PBS and collected using Accutase (Nacalai Tesque) and resuspended in 50 μL staining buffer (BD Pharmingen, country). The harvested cells were blocked with 2 μL Human Trustain FcX (Fc receptor Blocking Solution; BioLegend) for 10 min at room temperature, and stained with 2 μL antibodies namely, FITC anti-human CD73, FITC anti-human CD90, Alexa Fluor® 488 anti-human CD105, FITC anti-human CD34, PE anti-human CD45, FITC anti-human CD11b, Alexa Fluor® 488 anti-human CD11b, PE anti-human CD206, APC anti-human CD206, PE anti-human CD86 and APC anti-human CD86 (BioLegend) in the dark for 30 min at 4 °C.
Nakao Y., Fukuda T., Zhang Q., Sanui T., Shinjo T., Kou X., Chen C., Liu D., Watanabe Y., Hayashi C., Yamato H., Yotsumoto K., Tanaka U., Taketomi T., Uchiumi T., Le A.D., Shi S, & Nishimura F. (2020). Exosomes from TNF-α-treated human gingiva-derived MSCs enhance M2 macrophage polarization and inhibit periodontal bone loss. Acta biomaterialia, 122, 306-324.
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4

Multiparametric Flow Cytometry Panel

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Alexa Fluor® 700 anti-human CD3, (clone: HIT3a), FITC-anti human TCRαβ (clone: IP26), FITC-anti-human TCRγδ (clone: B1), APC/Cy7 Anti-Human CD127 (I-7Ra), (clone: A019D5), PE anti-human CD161, (clone: HP-3G10), Brilliant Violet 421TM anti-human CD117 (c-kit), (clone 104D2), PE/Cy7 anti-human CD294 (CRTH2), (clone: BM16), APC anti-human CD336 (NKp44), (clone: 325110), Alexa Fluor® 488 anti-human CD19, (clone: HIB19), FITC anti-human CD94, (clone:DX22), FITC anti-human CD1a, (clone: HI149), FITC anti-human CD11c, (clone: 3.9), FITC anti-human CD123, (clone: 6H6), anti-human CD303 (BDCA-2), (clone:201A), FITC anti-human CD14, (clone: 63D3), FITC anti-human FcεRIα, (clone: NP4D6), FITC anti-human CD34,(clone: 561), APC/Cy7 anti-human IFN-γ, (clone: 4S.B3) all from BioLegend. Anti-Human CD363 (S1PR1) eFluor® 660, (clone: SW4GYPP, ThermoFisher), Mouse IgG1 K Isotype Control eFluor® 660, (clone: P3.6.2.8.1, ThermoFisher). Anti-mouse CD3) APC, (clone: 17A2), anti-mouse NK1.1 Alexa Fluor® 647, (clone: PK136), anti-mouse B220 APC/Cy7 or FITC, (clone:RA3-6B2), anti-mouse CD45-Percp Cy5.5, (clone:30-F11), anti-mouse CD90.2 PE Cy7, (clone:30-H12), anti-mouse CD11b APC, (clone:M1/70), anti-mouse GATA3 PE (Clone: 16E10A23), anti-mouse Rorγt (Clone: 2F7-2).
Eken A., Yetkin M.F., Vural A., Okus F.Z., Erdem S., Azizoglu Z.B., Haliloglu Y., Cakir M., Turkoglu E.M., Kilic O., Kara I., Dönmez Altuntaş H., Oukka M., Kutuk M.S., Mirza M, & Canatan H. (2019). Fingolimod Alters Tissue Distribution and Cytokine Production of Human and Murine Innate Lymphoid Cells. Frontiers in Immunology, 10, 217.
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5

Flow Cytometric Characterization of hADSCs

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The hADSCs of passages 3 or 5 were collected with 1×Tryple Express and centrifuged at 400 g for 5 min. After washing twice with 1×DPBS, cells were resuspended and incubated with pre-labelled antibodies for 15 min at room temperature. After two washes with 1×PBS, cells were resuspended in 300 μL 1×PBS and analyzed using a flow cytometer (BD FACSCalibur). Histograms were generated using the CELLQuest Pro software (BD Biosciences). The antibodies used were as follows: FITC Mouse IgG1,k,Iso-type Ctrl (FC) (BioLegend, 400110), FITC anti-human CD34 (BioLegend, 343504), FITC anti-human CD45, (BioLegend, 304006), FITC anti-human CD11b (BioLegend, 301330), FITC anti-human HLA-DR (BioLegend, 307604), FITC anti-human CD73 (BioLegend, 344016), FITC anti-human CD90 (BioLegend, 328108), APC Mouse IgG1,k,Isotype Ctrl (BioLegend, 400120), APC anti-human CD19 (BioLegend, 363006), PE Mouse IgG1,k,Isotype Ctrl (FC) (BioLegend, 400114), PE anti-CD105 (Endoglin) (BioLegend, 800504).
Yan K., Zhang J., Yin W., Harding J.N., Ma F., Wu D., Deng H., Han P., Li R., Peng H., Song X, & Kang Y.J. (2022). Transcriptomic heterogeneity of cultured ADSCs corresponds to embolic risk in the host. iScience, 25(8), 104822.
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6

Multicolor Flow Cytometric Analysis of HSPCs

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HSPCs sorted using the microfluidic device were stained with FITC anti-human CD34, Alexa Fluor® 647 anti-human CD38, Brilliant Violet 421™ anti-human CD90, and PE/Cy7 anti-human CD45RA antibodies (BioLegend). Stained HSPCs were analyzed using a BD LSRFortessa cell analyzer.
Han X., Ma Y., Zhang K., Zhang P., Shao N, & Qin L. (2020). Microfluidic Cell Trap Arrays for Single Hematopoietic Stem/ Progenitor Cell Behavior Analysis. Proteomics, 20(13), e1900223.
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7

Flow Cytometry Analysis of Hematopoietic Stem Cells

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Cultured cells were collected at different time and incubated with the indicated antibodies for 30 min at 4 °C in PBS containing 0.5% BSA (Sigma, Cat: A1470–100G). Next, the cells were washed three times with PBS and suspended in 0.2 ml PBS for analysis. Flow cytometry analysis was performed using FACSVerse (BD) or LSRFortessa (BD). The data were analyzed using FlowJo-V10 (BD). The following antibodies were used: FITC anti-human CD34 (Biolegend, Cat: 343604), PE anti-human CD38 (Biolegend, Cat: 356604), PE-Cy7 anti-human CD49f (Biolegend, Cat: 313622), APC anti-human CD90 (Biolegend, Cat: 328114), APC-Cy7 anti-human CD45RA (Biolegend, Cat: 304128). For analysis of engrafted human hematopoietic lineages, peripheral blood cells were collected at indicated time. BM was isolated at 21 weeks post transplantation, and cells were stained with following antibodies: PE-Cy7 anti-human CD45 (Biolegend, Cat: 304016), FITC anti-mouse CD45 (Biolegend, Cat: 103108), PE anti-human CD19 (Biolegend, Cat: 302208), APC anti-human CD33 (Biolegend, Cat: 366606).
Xiao X., Lai W., Xie H., Liu Y., Guo W., Liu Y., Li Y., Li Y., Zhang J., Chen W., Shi M., Shang L., Yin M., Wang C, & Deng H. (2019). Targeting JNK pathway promotes human hematopoietic stem cell expansion. Cell Discovery, 5, 2.
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8

Immunophenotyping of GMSCs and Macrophages

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The expression of CD73, CD90, CD105, CD34, CD45, and CD11b in GMSCs and CD11b, CD206, and CD86 in PBMC-derived macrophages were analyzed using the FACS Calibur (Becton Dickinson, USA) and the CellQuest software (Becton Dickinson). The adherent cells were washed with PBS and collected using Accutase (Nacalai Tesque) and resuspended in 50 μL staining buffer (BD Pharmingen, country). The harvested cells were blocked with 2 μL Human Trustain FcX (Fc receptor Blocking Solution; BioLegend) for 10 min at room temperature, and stained with 2 μL antibodies namely, FITC anti-human CD73, FITC anti-human CD90, Alexa Fluor® 488 anti-human CD105, FITC anti-human CD34, PE anti-human CD45, FITC anti-human CD11b, Alexa Fluor® 488 anti-human CD11b, PE anti-human CD206, APC anti-human CD206, PE anti-human CD86 and APC anti-human CD86 (BioLegend) in the dark for 30 min at 4 °C.
Nakao Y., Fukuda T., Zhang Q., Sanui T., Shinjo T., Kou X., Chen C., Liu D., Watanabe Y., Hayashi C., Yamato H., Yotsumoto K., Tanaka U., Taketomi T., Uchiumi T., Le A.D., Shi S, & Nishimura F. (2020). Exosomes from TNF-α-treated human gingiva-derived MSCs enhance M2 macrophage polarization and inhibit periodontal bone loss. Acta biomaterialia, 122, 306-324.
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9

Isolation and Characterization of CD34+ Cells from Cord Blood and CML Patients

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Details on the cell lines used in this study are available in Supplementary Methods. Primary mononuclear cells (MNCs) from discarded cord blood from the University Medical Center of El Paso (El Paso, TX, USA) were Ficoll-separated (Life Technologies, Carlsbad, CA, USA) and used for selection of the CD34+ population (StemCell Technologies Inc., Vancouver, Canada). CD34+ cells represent the disease-causing stem and progenitor population in CML. A purity of >90% was confirmed by flow cytometry using FITC anti-human CD34 (BioLegend Inc., San Diego, CA, USA). CD34+ cells from peripheral blood of CML patients or normal donors that were used for nucleocytoplasmic fractionation (see below) were obtained from the Centre for Haematology at Imperial College London (London, UK). Cells (2 × 106) were lysed directly for use in molecular biology assays. This study was approved by the Texas Tech University Health Sciences Center El Paso Institutional Review Board (IRB) and by the Imperial College Research Ethics Committee. All patients gave informed consent and all experiments were performed in accordance with the Declaration of Helsinki.
Bencomo-Alvarez A.E., Rubio A.J., Olivas I.M., Gonzalez M.A., Ellwood R., Fiol C.R., Eide C.A., Lara J.J., Barreto-Vargas C., Jave-Suarez L.F., Nteliopoulos G., Reid A.G., Milojkovic D., Druker B.J., Apperley J., Khorashad J.S, & Eiring A.M. (2021). Proteasome 26S subunit, non-ATPases 1 (PSMD1) and 3 (PSMD3), play an oncogenic role in chronic myeloid leukemia by stabilizing nuclear factor-kappa B. Oncogene, 40(15), 2697-2710.
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10

Flow Cytometry Analysis of PDLSCs

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The expression of typical surface markers in PDLSCs at the third passage was analyzed using a Novocyte Flow Cytometer (ACEA Biosciences, San Diego, CA, USA). The adherent cells were collected and resuspended in 50 μL of phosphate-buffered saline (PBS). The harvested cells were stained with 5 μL antibodies, namely FITC anti-human CD31 (#303103, BioLegend, San Diego, CA, USA), FITC anti-human CD34 (#343603, BioLegend), FITC anti-human CD73 (#344016, BioLegend), FITC anti-human CD90 (#328107, BioLegend), or FITC anti-human CD146 (#361011, BioLegend), in the dark for 30 min at 4 °C. Raw data were analyzed using the FlowJo software (FlowJo, Ashland, OR, USA).
Lu Y., Zhao L., Mao J., Liu W., Ma W, & Zhao B. (2023). Rab27a-mediated extracellular vesicle secretion contributes to osteogenesis in periodontal ligament-bone niche communication. Scientific Reports, 13, 8479.
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