Human cd34 positive selection kit

Manufactured by STEMCELL

Sourced in Canada

The Human CD34 Positive Selection Kit is a laboratory tool designed for the isolation and enrichment of CD34-positive cells from human samples. It utilizes magnetic beads coated with anti-CD34 antibodies to selectively capture and separate the target cells from the rest of the cell population.

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Lab products found in correlation

Interleukin 3 (il 3), by Thermo Fisher Scientific (2 mentions) Stem cell factor (scf), by Thermo Fisher Scientific (2 mentions) Ficoll hypaque density gradient, by Cytiva (1 mentions) Rpmi 1640, by Merck Group (1 mentions) Thrombopoietin, by Thermo Fisher Scientific (1 mentions) Fms related tyrosine kinase 3 ligand, by Thermo Fisher Scientific (1 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions) Iscove modified dulbecco media (imdm), by GE Healthcare (1 mentions) G csf, by Thermo Fisher Scientific (1 mentions) Flt3l, by Thermo Fisher Scientific (1 mentions) Ficoll paque plus, by GE Healthcare (1 mentions) Interleukin 6 (il 6), by Thermo Fisher Scientific (1 mentions) Lymphoprep, by Boster Bio (1 mentions) Flt3 ligand, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

EasySep human whole blood CD34 positive selection kit EasySep Human CD34 Positive Selection Kit II EasySep™ Human Cord Blood CD34 Positive Selection Kit II EasySep Human CD34 Positive Selection Kit EasySep Human Cord Blood CD34 Positive Selection Kit Human Cord Blood CD34 Positive Selection Kit II CD34 positive selection kit

9 protocols using human cd34 positive selection kit

Isolation and Culture of K562 Cells and Primary CML CD34+ Cells

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The human myelogenous leukaemia cell line K562 were obtained from America Type Culture Collection (ATCC, Manassas, VA, USA) and cultured in RPMI-1640 (Sigma, St. Louis, MO, USA) medium containing 10% heat-inactivated fetal calf serum (FCS, Hyclone, Logan, UT, USA), 100 unit/ml penicillin and 100 μg/ml streptomycin in a humidified 5% CO₂ atmosphere at 37°C.
The bone marrow (BM) samples were obtained from healthy donor or CML patients undergoing diagnostic procedures at Peking university first hospital. Written informed consent was obtained from each healthy donor and CML patient. All the procedures were approved by the Ethics Committee of Beijing Institute of Radiation Medicine. Mononuclear cells were isolated from heparinized samples by centrifugation through a Ficoll-Hypaque density gradient (Amersham Biosciences, Piscataway, NJ, USA). Then, CD34⁺ cells were isolated by using human CD34 positive selection kit (Stem Cell Technology, Vancouver BC, Canada).

Yang Y., Liu X., Xiao F., Xue S., Xu Q., Yin Y., Sun H., Xu J., Wang H., Zhang Q., Wang H, & Wang L. (2015). Spred2 Modulates the Erythroid Differentiation Induced by Imatinib in Chronic Myeloid Leukemia Cells. PLoS ONE, 10(2), e0117573.

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Humanized Immune System Mouse Model

Cited 2 times

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Recombinant AAV9 (rAAV9) vectors encoding human IL-3, IL-15, GM-CSF, and HLA-A∗0201 were constructed as previously described [27] (link). Four-week-old NSG mice were transduced with rAAV9 encoding HLA-A∗0201 by intrathoracic (IT) injection and with rAAV9 encoding HLA-A∗0201 and AAV9 encoding human IL-3, IL-15, and GM-CSF, by intravenous (IV) injection, as previously described [27] (link). Two weeks later, mice were subjected to 150-Gy total body sub-lethal irradiation for myeloablation, and several hours later, each transduced, irradiated mouse was engrafted intravenously with 1 × 10⁵ HLA-A∗0201+ matched, CD34+ human hematopoietic stem cells (HSCs). CD34+ HSCs among lymphocytes derived from HLA-A∗0201+ fetal liver samples were isolated using a Human CD34 Positive Selection kit (STEMCELL TECHNOLOGIES Inc. Vancouver, BC, Canada) [28] (link). At 14 weeks after HSC engraftment, the reconstitution status of human CD45+ cells in the blood of HIS-CD8 mice was determined by flow cytometric analysis, as previously described [27] (link).

Li X., Huang J., Zhang M., Funakoshi R., Sheetij D., Spaccapelo R., Crisanti A., Nussenzweig V., Nussenzweig R.S, & Tsuji M. (2016). Human CD8+ T cells mediate protective immunity induced by a human malaria vaccine in human immune system mice. Vaccine, 34(38), 4501-4506.

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Engraftment of CML Stem/Progenitor Cells in NSG Mice

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Peripheral blood from CML patients were layered on Ficoll-Paque PLUS (1.077 g/ml) for gradient centrifugation. The layer of mononuclear cells was collected, washed, and enriched for CD34⁺ CML stem/progenitor cells by positive selection using Human CD34 Positive Selection Kit (StemCell Technologies). Depending on the numbers of recovered cells, 7×10⁴ to 1×10⁶ CD34⁺ CML stem/progenitor cells were transplanted into sub-lethally irradiated (2.25–2.5 Gy) NSG mice via tail-vein injection. The NSG recipients were treated with DMSO, PGE1 and/or imatinib, monitored and analyzed for engraftment of human CD45⁺ cells.

Li F., He B., Ma X., Yu S., Roy R., Lentz S.R., Tan K., Guzman M.L., Zhao C, & Xue H.H. (2017). Prostaglandin E1 and its analog misoprostol inhibit human CML stem cell self-renewal via EP4 receptor activation and repression of AP-1. Cell stem cell, 21(3), 359-373.e5.

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Engraftment of Transduced Human HSCs

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Lymphocytes were isolated from fetal liver samples as described previously [26 (link)]. CD34+ human HSCs were isolated from lymphocytes using the Human CD34 Positive Selection Kit (STEMCELL Technologies Inc., Vancouver, BC, Canada), according to the manufacturer’s instructions. HSC purity was evaluated using flow cytometric analysis, and the percentage of CD34+ cells was confirmed to be higher than 90%. One to two weeks after the transduction of selected human genes by AAV9 vectors, mice were exposed to 150-Gy whole-body sub-lethal irradiation for myeloablation. A few hours later, each transduced, irradiated mouse was engrafted IV with 1 × 10⁵ HLA-A*0201+ matched, CD34+ human HSCs.

Li X., Huang J., Kaneko I., Zhang M., Iwanaga S., Yuda M, & Tsuji M. (2016). A potent adjuvant effect of a CD1d-binding NKT cell ligand in human immune system mice. Expert review of vaccines, 16(1), 73-80.

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Erythroid Differentiation from CD34+ Cells

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To generate erythroid cells from co-cultured cells, co-D12 or co-D14 CD34⁺ cells were isolated using a human CD34 positive selection kit (Stem Cell Technologies). CD34⁺ cells (1 × 10⁵/ml) were suspended in serum-free expansion medium II (Stem Cell Technologies) with 50 ng/ml stem cell factor (PeproTech), thrombopoietin (PeproTech), Fms-related tyrosine kinase 3 ligand (PeproTech), and 0.5% penicillin/streptomycin (Invitrogen). After 3 days in HSPC expansion culture, the cells were cultured using a three-phase liquid culture system (described in Supplementary Methods). To generate UCB-derived erythroid cells, CD34⁺ cells were sorted using the same method described above, and then the cells were cultured in a three-phase liquid culture system.

Chen Y., Dong Y., Lu X., Li W., Zhang Y., Mao B., Pan X., Li X., Zhou Y., An Q., Xie F., Wang S., Xue Y., Cai X., Lai M., Zhou Q., Yan Y., Fu R., Wang H., Nakahata T., An X., Shi L., Zhang Y, & Ma F. (2022). Inhibition of aryl hydrocarbon receptor signaling promotes the terminal differentiation of human erythroblasts. Journal of Molecular Cell Biology, 14(2), mjac001.

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Autophagic Analysis of Human Blood Samples

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The clinical records in human blood test were provided by the Department of Orthopaedics, the Second Affiliated Hospital of Soochow University. Human blood samples for autophagic analysis were obtained from volunteer donors with their consent. All experiments with human samples and clinical records were approved by the Research Management Office of the Affiliated Hospital of Soochow University. Human mononuclear cells were enriched by equilibrium centrifugation over a cushion of Ficoll‐Paque Plus (17544652, GE Healthcare) by density gradient centrifugation. Human HSPCs were purified from bone marrow CD45‐positive cells by human CD34‐positive selection kit (18056, STEMCELL Technologies). Human HSPCs were cultured in IMDM (SH30228, GE Healthcare) with 10% FBS, 1% penicillin/streptomycin, SCF (100 ng/ml), Flt3L (100 ng/ml), IL6 (20 ng/ml), IL3 (20 ng/ml), and G‐CSF (20 ng/ml) (PeproTech).

Fang Y., An N., Zhu L., Gu Y., Qian J., Jiang G., Zhao R., Wei W., Xu L., Zhang G., Yao X., Yuan N., Zhang S., Zhao Y, & Wang J. (2020). Autophagy‐Sirt3 axis decelerates hematopoietic aging. Aging Cell, 19(10), e13232.

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Humanized Mouse Model Generation

Cited 2 times

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Recombinant AAV9 (rAAV9) vectors encoding human IL-3, IL-15, GM-CSF, and HLA-A*0201 were constructed as previously described (Huang et al., 2014 (link)). Four-week-old NSG mice were transduced with rAAV9 encoding HLA-A*0201 by perithoracic injection and with rAAV9-encoding HLA-A*0201 and AAV9-encoding human IL-3, IL-15, and GM-CSF, by IV injection, as previously described (Huang et al., 2014 (link)). Two weeks later, mice were subjected to 150-Gy total body sub-lethal irradiation for myeloablation, and several hours later, each transduced, irradiated mouse was engrafted IV with 1 × 10⁵ HLA-A*0201⁺-matched, CD34⁺ human HSCs. CD34⁺ HSCs among lymphocytes derived from HLA-A*0201⁺ fetal liver samples were isolated using a Human CD34 Positive Selection Kit (STEMCELL Technologies, Vancouver, BC, United States) (Lepus et al., 2009 (link)). At 14 weeks post-HSC engraftment, the reconstitution status of human CD45⁺ cells in the blood of HIS-CD8 mice was determined by flow cytometry analysis, as previously described (Huang et al., 2014 (link)).

Phares T.W., Huang J., Kotraiah V., Hauser M.J., Domi A., Oruganti S., Browne C.D., Buontempo P., Mansour M., Pannucci J., Tsuji M, & Gutierrez G.M. (2022). Viral delivery of a peptide-based immunomodulator enhances T cell priming during vaccination. Frontiers in Pharmacology, 13, 1029636.

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Isolation of CD34+ Cells from Bone Marrow

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CD34 + cells were isolated by Human CD34 Positive Selection Kit (StemCell Technologies, Vancouver, Canada) from BMMNCs according to manufacturer’s protocol. CD34+ cells purity was evaluated with Fluorescence Activated Cell Sorting (FACS) (BD Biosciences, Franklin Lakes, NJ, USA) and was >90%.

Zhao S., Zhao Y., Guo J., Fei C., Zheng Q., Li X, & Chang C. (2017). Downregulation of MMP1 in MDS-derived mesenchymal stromal cells reduces the capacity to restrict MDS cell proliferation. Scientific Reports, 7, 43849.

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CD34+ HSCs Expansion and Culture

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UCB mononuclear cells were separated by density gradient centrifugation with lymphoprep (Boster Bio, China), and then CD34⁺ HSCs were purified from the mononuclear cell fraction by using a human CD34-positive selection kit (Stem Cell Technology, Vancouver, Canada) according to the manufacturer’s protocol. For expansion of CD34⁺ cells, hematopoietic growth factors were added as follows: IL-3 10 ng/mL, SCF 100 ng/mL, TPO 100 ng/mL, Flt-3 ligand 100 ng/mL (PeproTech, USA). About 4×10⁴ cells were seeded in each plate well. The cells were cultured at 37°C in a humidified atmosphere of 5% CO₂ and harvested on Day 0, 6, and 12. Cell pellets and supernatants were stored at −80°C for further study.

Huang Y., Yan Q., Fan R., Song S., Ren H., Li Y, & Lan Y. (2016). Hepatitis B Virus Replication in CD34+ Hematopoietic Stem Cells From Umbilical Cord Blood. Medical Science Monitor : International Medical Journal of Experimental and Clinical Research, 22, 1673-1681.

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