Monocyte enrichment cocktail

Manufactured by STEMCELL

Sourced in Canada

The Monocyte Enrichment Cocktail is a cell separation product designed to isolate monocytes from human peripheral blood mononuclear cells (PBMCs). The cocktail contains a combination of monoclonal antibodies and magnetic particles that selectively bind to non-monocytic cells, allowing for the efficient separation and enrichment of monocytes.

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Lab products found in correlation

Moflo xdp, by Beckman Coulter (1 mentions) Cd16 fitc clone 3g8, by BD (1 mentions) Cd14 rd1 clone 322a 1, by Beckman Coulter (1 mentions) Rpmi 1640, by Thermo Fisher Scientific (1 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions) Lymphoprep, by STEMCELL (1 mentions) Sepmate 50, by STEMCELL (1 mentions) Facsaria 2 cell sorter, by BD (1 mentions) Facscelesta, by BD (1 mentions)

Close spelling variants of this product

RosetteSep Human Monocytes enrichment cocktail RosetteSep™ human monocyte enrichment cocktail kit RosetteSep Human monocyte Cell Enrichment Cocktail Rosette Sep® monocyte enrichment cocktail RosetteSep Human Monocyte Enrichment Cocktail Human monocyte enrichment cocktail

4 protocols using monocyte enrichment cocktail

Monocyte Subsets Isolation and Stimulation

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MDM from HD and patients with RA were obtained as explained before. Monocytes were isolated from 50 mL of venous blood from HD using Monocyte Enrichment Cocktail (Stem Cell Technologies) following the manufacturer’s instructions. Total monocytes (purity > 90%) were stained with CD14-RD1 (clone 322A-1; Beckman Coulter, CA) and CD16-FITC (clone 3G8; BD) antibodies for 15 min at 4 °C. CD16 − (purity > 95%) and CD16 + (purity > 90%) monocytes were sorted by using MoFlo XDP (Beckman Coulter, CA).
MDM were cultured for 6 h in the absence or presence of PRA-m/lEVs or PRA-m/lEV-ICs at a ratio monocyte:m/lEVs of 1:3. Enriched monocytes were cultured for 24 h as previously described [34 (link)], in the absence or presence of PRA-m/lEVs or PRA-m/lEV-ICs at ratio monocyte:m/lEVs of 1:3. Supernatants were stored at − 20 °C.

Rincón-Arévalo H., Burbano C., Atehortúa L., Rojas M., Vanegas-García A., Vásquez G, & Castaño D. (2022). Modulation of B cell activation by extracellular vesicles and potential alteration of this pathway in patients with rheumatoid arthritis. Arthritis Research & Therapy, 24, 169.

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Generation of Polarized Human Macrophages

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HPBMo were obtained using a monocyte enrichment cocktail (StemCell Technologies, Vancouver, BC, Canada) according to the manufacturer's instructions, seeded on Repcell dishes (CellSeed, Tokyo, Japan) for flow cytometric analysis or 12-well plates for the other analyses, and differentiated into HMDM by culturing for 8–11 days in RPMI 1640 (Thermo Fisher Scientific, Waltham, MA) containing 10% human serum, 1% penicillin-streptomycin, and 10 ng/ml human M-CSF (RPMI/M-CSF). Then, the HMDM were either maintained for 6 days in RPMI/M-CSF (nonpolarized HMDM), or cultured in RPMI/M-CSF containing 40 ng/ml human IFN-γ, 20 ng/ml IL-4, or 40 ng/ml IL-10 to elicit polarization into M1, M2a, and M2c subsets, respectively. The cells were cultured at 37 °C in an atmosphere of 5% CO₂ in air at 95% humidity. The resulting cells were used to prepare RNA and cell lysate, and for immunocytochemistry and flow cytometric analysis as described in the Supporting information.

Hayashi H., Naoi S., Togawa T., Hirose Y., Kondou H., Hasegawa Y., Abukawa D., Sasaki M., Muroya K., Watanabe S., Nakano S., Minowa K., Inui A., Fukuda A., Kasahara M., Nagasaka H., Bessho K., Suzuki M, & Kusuhara H. (2017). Assessment of ATP8B1 Deficiency in Pediatric Patients With Cholestasis Using Peripheral Blood Monocyte-Derived Macrophages. EBioMedicine, 27, 187-199.

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Monocyte-Derived Macrophage Generation and HIV-1 Production

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Monocytes were isolated from buffy coat from healthy blood donors by positive selection on Monocyte Enrichment Cocktail (Stemcell, Tukwila, WA) and Lymphoprep (Stemcell) density gradient centrifugation with SepMate-50 (Stemcell). MDMs were generated by culturing monocytes with 100 ng/ml granulocyte-macrophage colony-stimulation factor for 5 days. 293T and HeLa cells were cultured in DMEM (Gibco-BRL, Rockville, MD) supplemented with 10% (V/V) fetal bovine serum (FBS) (Gibco-BRL). HIV-1_89.6 and HIV-1_NLAD-8 strains were produced in 293T cells. Vesicular stomatitis virus G glycoprotein (VSV-G)-pseudotyped viruses were produced in 293T cells cotransfected with reporter virus plasmid and VSV-G using the calcium-phosphate method. The culture supernatants were collected and subjected to quantification of HIV-1 particle yields by p24CA antigen capture enzyme-linked immunosorbent assay (ELISA) (Zepto Metrix, Buffalo, NY). Monocyte isolation and treatment were approved by the Ethics Committee at the Yokohama City University School of Medicine.

Kudoh A., Takahama S., Sawasaki T., Ode H., Yokoyama M., Okayama A., Ishikawa A., Miyakawa K., Matsunaga S., Kimura H., Sugiura W., Sato H., Hirano H., Ohno S., Yamamoto N, & Ryo A. (2014). The phosphorylation of HIV-1 Gag by atypical protein kinase C facilitates viral infectivity by promoting Vpr incorporation into virions. Retrovirology, 11, 9.

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Isolation and Polarization of Human Peripheral Blood Monocytes

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Peripheral blood samples from patients with PFIC1 or BRIC1 and age‐matched control subjects, including healthy individuals and those with pancreatitis, hepatitis B and C virus, and biliary atresia, were collected in ethylene diamine tetraacetic acid‐2K‐coated blood sampling tubes (Becton Dickinson, Tokyo, Japan). Human peripheral blood monocytes (HPBMo) were obtained using a monocyte enrichment cocktail (StemCell Technologies, Vancouver, Canada), seeded on Repcell dishes (CellSeed, Tokyo, Japan), and cultured at 37°C in an atmosphere of 5% CO₂ in air at 95% humidity. HPBMo from more than 3 age‐matched control subjects were pooled before the seeding and then employed as control cells for flow cytometric analysis. HPBMo were differentiated into HMDMs and then polarized into M2c subsets, as described.⁽²⁸⁾ The prepared cells were harvested by incubating the dishes on ice, stained with fluorochrome‐labeled antibodies, and analyzed on a BD FACSAria II Cell Sorter (BD Biosciences, San Jose, CA) or BD FACSCelesta (BD Biosciences).

Mizutani A., Sabu Y., Naoi S., Ito S., Nakano S., Minowa K., Mizuochi T., Ito K., Abukawa D., Kaji S., Sasaki M., Muroya K., Azuma Y., Watanabe S., Oya Y., Inomata Y., Fukuda A., Kasahara M., Inui A., Takikawa H., Kusuhara H., Bessho K., Suzuki M., Togawa T, & Hayashi H. (2020). Assessment of Adenosine Triphosphatase Phospholipid Transporting 8B1 (ATP8B1) Function in Patients With Cholestasis With ATP8B1 Deficiency by Using Peripheral Blood Monocyte‐Derived Macrophages. Hepatology Communications, 5(1), 52-62.

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