Human thymus samples were collected from pediatric patients undergoing cardiac surgery (under the clinical protocol TIGET07, approved by the Ethical Committee of San Raffaele Scientific Institute). Thymic tissue was cleaned and minced into small pieces before digestion with Liberase/DNase I (Sigma-Aldrich). Digested tissue was depleted of CD45-positive cells using the AutoMACS Pro separator (Miltenyi Biotec) after incubation with anti–human CD45 microbeads (Miltenyi Biotec). After depletion, thymic cells were stained with antibodies anti–human CD45 APC, epithelial cell adhesion molecule/CD326 Pe-Vio770, HLADR PE, CD31 APC-Vio770 (all from Miltenyi Biotec), and Ulex europaeus agglutinin I FITC (UEA I FITC; Vector Laboratories). After surface staining, thymic cells were fixed/permeabilized (Foxp3 Staining Buffer set; eBioscience) and stained intracellularly with anti-Extl3 antibody (Ab113063; Abcam), followed by a secondary goat anti–rabbit IgG Alexa Fluor 405 antibody (Invitrogen). Then, cells were acquired on a FACS Canto II cell analyzer (BD) and analyzed using FlowJo software (Tree Star).
Anti human cd45 apc
Manufactured by Miltenyi Biotec
Sourced in Germany
Anti-human CD45 APC is a monoclonal antibody that binds to the CD45 antigen expressed on the surface of human leukocytes. The APC (Allophycocyanin) fluorochrome is conjugated to the antibody, allowing for the detection and identification of CD45-positive cells by flow cytometry.
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Lab products found in correlation
Anti human cd45 microbeads, by Miltenyi Biotec (1 mentions)
Liberase dnase 1, by Merck Group (1 mentions)
Hla dr pe, by Miltenyi Biotec (1 mentions)
Facscanto 2 cell analyzer, by BD (1 mentions)
Foxp3 staining buffer set, by Thermo Fisher Scientific (1 mentions)
Automacs pro separator, by Miltenyi Biotec (1 mentions)
Anti human cd34 fitc, by Miltenyi Biotec (1 mentions)
Cytoflex, by Beckman Coulter (1 mentions)
Cytexpert software, by Beckman Coulter (1 mentions)
2 protocols using anti human cd45 apc
1
Isolation and Characterization of Human Thymic Cells
2
Phenotyping Hematopoietic Progenitor Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
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To compare the efficiency of differentiation into HPC, HPC-EB was dissociated with Accutase for 10 min at 37 °C in 5% CO2 incubator and then gently pipetted into single cells. After the dissociated cells were centrifuged for 5 min, the supernatant was aspirated and resuspended in the FACS buffer. The cells were then incubated with fluorescence-conjugated primary antibodies, anti-human CD34-FITC (1:100, Miltenyi Biotec, Germany), and anti-human CD45-APC (1:100, Miltenyi Biotec, Germany) for 30 min on ice. After three washes with FACS buffer, the fluorescence signal was measured using Beckman Coulter/CytoFLEX (Beckman Coulter, Indianapolis, IN, USA) and examined using CytExpert software (Beckman Coulter).
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