Cd1a microbeads

Manufactured by Miltenyi Biotec

Sourced in United States

CD1a microbeads are a lab equipment product used for the isolation and enrichment of CD1a-positive cells. They provide a tool for the study and investigation of this specific cell population.

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Lab products found in correlation

Recombinant human il 4, by R&D Systems (2 mentions) Recombinant human gm csf, by R&D Systems (2 mentions) 40 m cell strainer, by BD (2 mentions) Automacs separator, by Miltenyi Biotec (2 mentions) Ficoll paque, by GE Healthcare (2 mentions) Collagenase d, by Roche (2 mentions) Ficoll gradient, by Miltenyi Biotec (2 mentions) Dispase 2, by Roche (2 mentions) Dispase, by Thermo Fisher Scientific (1 mentions) Rpmi 10 fcs, by Thermo Fisher Scientific (1 mentions) Rpmi 10 fcs, by Biowest (1 mentions) Dnase 1, by Roche (1 mentions) Collagenase, by Worthington (1 mentions) Gm csf, by Thermo Fisher Scientific (1 mentions) Bovine serum albumin (bsa), by Merck Group (1 mentions) Rpmi 1640, by Lonza (1 mentions) Cd14 microbead, by Miltenyi Biotec (1 mentions)

7 protocols using cd1a microbeads

Isolation and Purification of Skin Cells

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For isolation of human skin cells 300 µm dermatome sections were incubated in RPMI+10%FCS (BioWest) containing 0.8 mg/ml collagenase (Type IV, Worthington-Biochemical) and 0.05 mg/ml DNase I (Roche) for 12 h. For nanostring analysis and T cell proliferation assay skin was treated with 1 mg/ml dispase (Invitrogen) to separate epidermis and dermis. Dermal DCs were sorted by fluorescence-activated cell sorting (FACS), epidermal LCs were isolated using CD1a microbeads (Miltenyi Biotec) and a magnet (Stemcell techonologies) with a purity of >90%.
For isolation of mouse skin cells, mice were sacrificed and ears were cut off at the base. Ear skin was split into dorsal and ventral halves and incubated in RPMI+10%FCS containing 1 mg/ml dispase (Invitrogen) for 2 h at 37deg. Epidermis and dermis were separated and digested in 0.2 mg/ml collagenase (Type IV, Sigma) for 2 h at 37deg before passing them through a 70 um filter to obtain a single cell suspension.
Mouse skin-draining auricular lymph nodes were isolated, incubated in medium+0.2 mg/ml collagenase for 30 min and passed through 70 um filter.
BHK-21 and C6/36 cells were purchased from the American Type Culture Collection.

Cerny D., Haniffa M., Shin A., Bigliardi P., Tan B.K., Lee B., Poidinger M., Tan E.Y., Ginhoux F, & Fink K. (2014). Selective Susceptibility of Human Skin Antigen Presenting Cells to Productive Dengue Virus Infection. PLoS Pathogens, 10(12), e1004548.

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Isolation of Human Lung Dendritic Cells

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Human lung single cell suspensions were prepared as previously described⁶. Briefly, fresh lung tissue was cut into 0.1 cm pieces in Petri dishes and treated with 2 mg/ml of collagenase D (Roche Pharmaceuticals) in HBSS for 30 to 40 min at 37 °C. Single cells were collected by mincing and pressing lung tissue through 40 µm cell strainers (BD Biosciences) followed by RBC lysis. Lung DCs were isolated by labeling RBC-free lung cells with CD1a Microbeads (Miltenyi Biotec) and then isolated by AutoMACS separator (Miltenyi). PBMCs were isolated by Ficoll-Paque (GE Healthcare) density gradient centrifugation. Human MDDCs were prepared as previously described. Briefly, RBC-free PBMCs were seeded in 6-well plates for two hours at 37°C and then non-adherent cells were removed by washing with PBS. Adherent cells were cultured with 50 ng/ml recombinant human GM-CSF and 10 ng/ml recombinant human IL-4 (R&D Systems Inc.) for 5 or 6 days. Both adherent and non-adherent cells were collected by gently scraping for further assays.

Lu W., You R., Yuan X., Yang T., Samuel E.L., Marcano D.C., Sikkema W.K., Tour J.M., Rodriguez A., Kheradmand F, & Corry D.B. (2015). MicroRNA-22 Inhibits Histone Deacetylase 4 to Promote T Helper-17 Cell-Dependent Emphysema. Nature immunology, 16(11), 1185-1194.

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Quantifying Cytosolic and Mitochondrial DNA in Dendritic Cells Co-cultured with Ovarian Cancer Cells

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Cytosolic DNA and mitochondrial DNA from PBMC-derived dendritic cells co-cultured with ovarian cancer cells were prepared as described (11 (link)), with modifications. In brief, dendritic cells were cultured in the top chamber of a modified transwell with ovarian cancer cells treated with RAMBO and/or cisplatin in the bottom chamber. Dendritic cells were harvested and purified with CD1a microbeads (130-051-001, Miltenyi Biotec,USA) and 1 × 10⁶ dendritic cells were divided into two parts with equal cell numbers (5 × 10⁵). One part was used to prepare total DNA as input. Another part was used to prepare cytosolic DNA by treating the cells with cytosolic extract buffer containing 150 mM NaCl, 50 mM HEPES and 25μg/ml digiton. Cytosolic DNA and mitochondrial DNA were quantified by qRT-PCR analysis of POLG expression (cytosolic DNA) and MT-ND1 expression (mitochondrial DNA). Primer sequences for PLOG and MT-ND1 genes are listed in Table 1.

Hong B., Chapa V., Saini U., Modgil P., Cohn D.E., He G., Siddik Z.H., Sood A.K., Yan Y., Karuppaiyah S., Pei G., Zhao Z., Yoo J.Y, & Kaur B. (2020). Oncolytic HSV therapy modulates vesicular trafficking inducing cisplatin sensitivity and anti-tumor immunity. Clinical cancer research : an official journal of the American Association for Cancer Research, 27(2), 542-553.

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Isolation of Vaginal Langerhans Cells

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Vaginal mucosa was obtained from routinely discarded tissue of vaginal prolapse surgeries. The study was approved by Medical Ethics Review Committee in accordance with the ethical guidelines of the Academic Medical Center. After incubation with Dispase II (3 mg/mL, Roche Diagnostics) in IMDM, vaginal mucosal sheets were separated from submucosa and further cultured in IMDM supplemented with 10% FCS, gentamycine (10 mg/mL), penicillin (2500 U/ml), streptomycin (2500 mg/ml), and L-Glutamine (100 mmol/l) until disintegration of the tissue. Further vaginal LC purification was performed using a Ficoll gradient and CD1a microbeads (Miltenyi Biotec).

Sarrami-Forooshani R., Mesman A.W., van Teijlingen N.H., Sprokholt J.K., van der Vlist M., Ribeiro C.M, & Geijtenbeek T.B. (2014). Human immature Langerhans cells restrict CXCR4-using HIV-1 transmission. Retrovirology, 11, 52.

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Isolation of Human Lung Dendritic Cells

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Vaginal Langerhans Cell Isolation and HIV Infection

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Vaginal mucosa was incubated with dispase II (3 mg/mL, Roche Diagnostics) in IMDM, to permit separation of mucosa from submucosa. Mucosal sheets were subsequently cultured in fully supplemented IMDM until the tissue disintegrated. Further vaginal LC purification was performed using a Ficoll gradient and CD1a microbeads (Miltenyi Biotec). Isolated LCs were routinely > 80% pure and expressed high levels of Langerin and CD1a. CD1a⁺ vaginal LCs were pre-incubated with 30 nM everolimus for 2 h before infection with HIV-1 NL4.3BaL at MOI = 0.09.

Cloherty A.P., van Teijlingen N.H., Eisden T.J., van Hamme J.L., Rader A.G., Geijtenbeek T.B., Schreurs R.R, & Ribeiro C.M. (2021). Autophagy-enhancing drugs limit mucosal HIV-1 acquisition and suppress viral replication ex vivo. Scientific Reports, 11, 4767.

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Generation of CD1a+ and CD14+ DCs

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CD34 + cells only from mobilized PB were cultured (10 5 cells/ml) in RPMI 1640 (BioWhittaker, Walkersville, MD, USA) containing 10% BSA (Sigma Aldrich, St. Louis, MO, USA) with L-glutamine and antibiotics (10%-RPMI), hereafter referred as medium, supplemented with 50 ng/ml of granulocyte macrophage-colony stimulating factor (GM-CSF; Peprotech, Rocky Hill, NJ, USA), 10 ng/ml of tumor necrosis factor (TNF)-α (Endogen, Rockford, IL, USA) and 50 ng/ml of Fms-related tyrosine kinase 3 ligand (FLT3L; eBioscience, San Diego, CA, USA) for 7 days [20] . CD34 + -derived DCs were harvested after 7 days of culture, and used for the purification of CD1a + cells by using CD1a microbeads (Miltenyi Biotec).
CD1a -fraction was used for the positive selection of CD1a -CD14 + cell population by using CD14 microbeads (Miltenyi Biotec).

Očadlíková D., Trabanelli S., Salvestrini V., Ciciarello M., Evangelisti C., Lecciso M., Sabattini E., Righi S., Piccioli M., Pileri S.A., Lemoli R.M, & Curti A. (2015). CD103 marks a subset of human CD34+-derived langerin+ dendritic cells that induce T-regulatory cells via indoleamine 2,3-dioxygenase-1. Experimental hematology, 43(4).

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