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Cd45 macs microbeads

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CD45+ MACS microbeads are magnetic particles coated with antibodies specific for the CD45 cell surface antigen. They are designed for the isolation and enrichment of CD45+ cells from complex samples, such as peripheral blood, bone marrow, or other tissues.

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Lab products found in correlation

Gentra puregene dna extraction kit, by Qiagen (2 mentions) Easysep mouse cd11b positive selection kit, by STEMCELL (1 mentions)

4 protocols using cd45 macs microbeads

1

Isolation of Colonic Cell Subsets

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Colonic cells were extracted as described above and subjected to cell debris depletion kit (Miltenyi Biotec) followed by column based magnetic cells separation. For LP and IE cells isolation, cells were subjected to EasySep™ Mouse CD11b positive selection kit (StemCell) and CD11b+ cells were collected. Negative fraction containing CD11b− cells were further subjected to CD45 MACS MicroBeads (Miltenyi Biotec) for CD11b− lymphocytes isolation.
Shmuel-Galia L., Humphries F., Lei X., Ceglia S., Wilson R., Jiang Z., Ketelut-Carneiro N., Foley S.E., Pechhold S., Houghton J., Muneeruddin K., Shaffer S.A., McCormick B.A., Reboldi A., Ward D., Marshak-Rothstein A, & Fitzgerald K.A. (2021). Dysbiosis exacerbates colitis by promoting ubiquitination and accumulation of the innate immune adaptor STING in myeloid cells. Immunity, 54(6), 1137-1153.e8.
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2

Inherited Platelet Disorder Genetics

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All patients in the original cohort (family 1) were recruited at the Hematology Clinic at Children's Hospital of Michigan. The study received Institutional Review Board approval from the University of Colorado Anschutz Medical Campus, and informed consent was obtained for all participants. Patients in families 2 and 3 were recruited in clinical centers in the Czech Republic and Italy as part of a European Consortium focused on inherited platelet disorders. The Institutional Review Board of the IRCCS Policlinico San Matteo Foundation of Pavia, Italy, approved the study protocol. Informed consent was obtained from all patients in the study. Studies were performed in accordance with the Declaration of Helsinki. Genomic DNA was extracted from whole blood using the Gentra Puregene DNA Extraction Kit (Qiagen). Platelets were purified from whole blood and negatively selected using CD45+ MACS microbeads (Miltenyi Biotec). Bone marrow aspirates were obtained for diagnosis and permission was obtained from families to use images.
Noetzli L., Lo R.W., Lee-Sherick A.B., Callaghan M., Noris P., Savoia A., Rajpurkar M., Jones K., Gowan K., Balduini C., Pecci A., Gnan C., De Rocco D., Doubek M., Li L., Lu L., Leung R., Landolt-Marticorena C., Hunger S., Heller P., Gutierrez-Hartmann A., Xiayuan L., Pluthero F.G., Rowley J.W., Weyrich A.S., Kahr W.H., Porter C.C, & Di Paola J. (2015). Germline mutations in ETV6 are associated with thrombocytopenia, red cell macrocytosis and predisposition to lymphoblastic leukemia. Nature genetics, 47(5), 535-538.
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3

Inherited Platelet Disorder Genetics

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All patients in the original cohort (family 1) were recruited at the Hematology Clinic at Children's Hospital of Michigan. The study received Institutional Review Board approval from the University of Colorado Anschutz Medical Campus, and informed consent was obtained for all participants. Patients in families 2 and 3 were recruited in clinical centers in the Czech Republic and Italy as part of a European Consortium focused on inherited platelet disorders. The Institutional Review Board of the IRCCS Policlinico San Matteo Foundation of Pavia, Italy, approved the study protocol. Informed consent was obtained from all patients in the study. Studies were performed in accordance with the Declaration of Helsinki. Genomic DNA was extracted from whole blood using the Gentra Puregene DNA Extraction Kit (Qiagen). Platelets were purified from whole blood and negatively selected using CD45+ MACS microbeads (Miltenyi Biotec). Bone marrow aspirates were obtained for diagnosis and permission was obtained from families to use images.
Noetzli L., Lo R.W., Lee-Sherick A.B., Callaghan M., Noris P., Savoia A., Rajpurkar M., Jones K., Gowan K., Balduini C., Pecci A., Gnan C., De Rocco D., Doubek M., Li L., Lu L., Leung R., Landolt-Marticorena C., Hunger S., Heller P., Gutierrez-Hartmann A., Xiayuan L., Pluthero F.G., Rowley J.W., Weyrich A.S., Kahr W.H., Porter C.C, & Di Paola J. (2015). Germline mutations in ETV6 are associated with thrombocytopenia, red cell macrocytosis and predisposition to lymphoblastic leukemia. Nature genetics, 47(5), 535-538.
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4

Isolation of Lung Leukocytes

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The left lung was minced in warmed digestion solution (RPMI 1640 with 5% FCS, 0.5 mg/ml Liberase TL and 40 U/ml DNase I). Digestion was stopped with cold HBSS containing 10 mM EDTA and the digested tissue was pushed through a 40 μm cell strainer and washed several times with wash buffer (RPMI 1640 with 5% FCS). Red blood cells were removed using ACK red blood cell lysis buffer (eBioscience, San Diego, CA). Lung digest was resuspended in wash buffer, and an aliquot was removed for cell count and viability using the Cellometer Auto 2000. Cells were centrifuged and resuspended in MACS/FACS buffer (HBSS w/o Ca and Mg, 2 mM EDTA and 0.5% BSA) containing CD45 MACS microbeads (Miltenyi Biotec, Bergisch Gladbach, Germany). CD45+ lung leukocytes were isolated using a Miltenyi LS column and magnetic separator.
Ogden H.L., Lai Y., Nolin J.D., An D., Frevert C.W., Gelb M.H., Altemeier W.A, & Hallstrand T.S. (2020). Secreted phospholipase A2 group X acts as an adjuvant for type-2 inflammation leading to an allergen-specific immune response in the lung. Journal of immunology (Baltimore, Md. : 1950), 204(12), 3097-3107.
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