Human monocyte isolation kit

Manufactured by Miltenyi Biotec

Sourced in United States

The Human Monocyte Isolation Kit is a laboratory equipment product designed to isolate human monocytes from whole blood or buffy coat samples. The kit utilizes a magnetic bead-based separation method to effectively separate and enrich the monocyte population from other blood cell types.

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Lab products found in correlation

Ammonium chloride potassium ack lysing buffer, by Thermo Fisher Scientific (1 mentions) Sepmate tube, by STEMCELL (1 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions) M csf, by Thermo Fisher Scientific (1 mentions) Ficoll paque, by GE Healthcare (1 mentions) Ficoll paque plus kit, by GE Healthcare (1 mentions) X vivo 15 serum free hematopoietic cell medium, by Lonza (1 mentions) Af 300 25, by Thermo Fisher Scientific (1 mentions)

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Monocyte Isolation Kit (79 citations) Isolation kits (10 citations)

4 protocols using human monocyte isolation kit

Isolation and Cryopreservation of Human Monocytes

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PBMCs were isolated using Sepmate tubes (StemCell Technologies) and ACK (ammonium-chloride-potassium) lysing buffer (Thermo Fisher Scientific) following the manufacturer’s protocol from buffy coats or leukopaks from healthy donors. Human monocytes were isolated from the PBMCs using the human monocyte isolation kit (Miltenyi) according to the manufacturer’s protocol. Monocytes were aliquoted and frozen in 10% DMSO and FBS for further use.

Freund E.C., Lock J.Y., Oh J., Maculins T., Delamarre L., Bohlen C.J., Haley B, & Murthy A. (2020). Efficient gene knockout in primary human and murine myeloid cells by non-viral delivery of CRISPR-Cas9. The Journal of Experimental Medicine, 217(7), e20191692.

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Isolation and Differentiation of Monocytes from Healthy Donors

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Peripheral blood mononuclear cells from de-identified healthy donors (below 35 years old) were obtained from the Stanford Blood Center. Samples were diluted with 20 mL PBS and layered onto 10 mL of Ficoll-Paque (GE Healthcare) using a pasteur pipette. Tubes were centrifuged at 1,500 rpm for 25 min without brake at 20°C. The mononuclear cell layer was transferred to a new 50 mL conical tube, resuspended in 50 mL 1× PBS, and centrifuged at 1,500 rpm for 10 min; this was repeated twice. After centrifugation, monocytes were isolated using a human Monocyte Isolation Kit, (Miltenyi Biotech). Cells were then plated 10 × 10⁶ per 10 cm dish and differentiated for 7 days in Roswell Park Memorial Institute (RPMI) medium supplemented with 10% FBS, 1% penicillin–streptomycin and 50 ng mL−1 M-CSF (Peprotech). Following differentiation, all experimental treatments including controls were done using RPMI containing 0.5% FBS (v/v).

Lin A.J., Joshi A.U., Mukherjee R., Tompkins C.A., Vijayan V., Mochly-Rosen D, & Haileselassie B. (2022). δPKC-Mediated Drp1 Phosphorylation Impacts Macrophage Mitochondrial Function and Inflammatory Response to Endotoxin. Shock (Augusta, Ga.), 57(3), 435-443.

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Isolation of Monocytes from Acute Gouty Arthritis

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At first, 30 mL of peripheral blood samples were gained from acute gouty arthritis patients and healthy volunteers. The Ficoll-Paque-PLUS kit (GE Healthcare, Piscataway, NJ, USA) was then used to isolate peripheral blood mononuclear cells (PBMCs). Subsequently, monocytes were isolated from PBMCs using the Human Monocyte Isolation Kit (Miltenyi Biotec, Auburn, CA, USA). The collected cells for enzyme-linked immunosorbent assay (ELISA) assay were stored at −80°C.

Zhao Q., Xia N., Xu J., Wang Y., Feng L., Su D, & Cheng Z. (2023). Pro-Inflammatory of PRDM1/SIRT2/NLRP3 Axis in Monosodium Urate-Induced Acute Gouty Arthritis. Journal of Innate Immunity, 15(1), 614-628.

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Bone Marrow and Monocyte-Derived Macrophage Isolation

Cited 1 time

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Bone marrow derived macrophages (BMDMs) were prepared as previously described (Colegio et al., 2014 (link)). For assaying Gdf15 expression in response to various stimuli, BMDMs were plated in either 48 well or 24 well plates at a density of 2.5×10⁵ or 5×10⁵ per well, respectively.
Human monocyte-derived macrophages (MDMs) were derived from human peripheral blood mononuclear cells (PBMCs). Initially, monocytes were isolated from cryopreserved PBMCs as per protocol from a human monocyte isolation kit (130-096-537, Miltenyi Biotec). Monocytes were then cultured at 2.5E6/ml in X-VIVO 15 serum-free hematopoietic cell medium (Lonza) supplemented with 50 ng/ml of human M-CSF (AF-300-25, Peprotech) for five days. Cells were then re-plated prior to cell treatments.

Eisenstein A., Hilliard B.K., Pope S.D., Zhang C., Taskar P., Waizman D.A., Israni-Winger K., Tian H., Luan H.H, & Wang A. (2022). Activation of the transcription factor NRF2 mediates the anti-inflammatory and tissue protective properties of a subset of over-the-counter and prescription NSAIDs. Immunity, 55(6), 1082-1095.e5.

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