Blood was layered on an equal volume of lymphocyte separation medium (Histopaque®-1077, Sigma-Aldrich, USA), followed by centrifugation for 30 min at 400g (room temperature). Subsequently, the peripheral blood monocytes were collected from the interface and washed with isotonic phosphate-buffered saline (PBS). The monocytes were isolated by a flow cytometer. Briefly, the monocytes were resuspended in 100 µl of PBS at 106 cells/sample. Then, 5 µl of mouse anti-human CD14 antibody (BD Pharmingen) was added. After incubation on ice for 30 min, the sample was washed three times with PBS at 4°C. The sample was resuspended in 500 µl of PBS and subjected to flow cytometry to isolate the monocytes.
Mouse anti human cd14 antibody
Manufactured by BD
Sourced in Sweden
The mouse anti-human CD14 antibody is a laboratory reagent used in immunological studies. It recognizes the CD14 cell surface antigen, which is primarily expressed on monocytes and macrophages. This antibody can be used to identify and isolate these cell types in various experimental applications.
Automatically generated - may contain errors
Lab products found in correlation
Histopaque 1077, by Merck Group (1 mentions)
Ham s f10, by Thermo Fisher Scientific (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Fetal calf serum (fcs), by Thermo Fisher Scientific (1 mentions)
Epidermal growth factor (egf), by Thermo Fisher Scientific (1 mentions)
Dexamethasone (dex), by Merck Group (1 mentions)
Insulin, by Merck Group (1 mentions)
Gentamycin, by Thermo Fisher Scientific (1 mentions)
Uridine, by Merck Group (1 mentions)
Pyruvate, by Merck Group (1 mentions)
Fetuin, by Merck Group (1 mentions)
Ficoll paque gradient separation, by GE Healthcare (1 mentions)
Creatine, by Merck Group (1 mentions)
2 protocols using mouse anti human cd14 antibody
1
Isolation of Peripheral Blood Monocytes
2
Isolation and Expansion of Human Skeletal Myoblasts
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Human skeletal muscle cells were isolated as previously described. 22 Freshly isolated cells were expanded in growth medium (GM) consisting of Ham's F10 (Life Technologies) supplemented with 15% FCS (Life Technologies), bovine serum albumin (Sigma-Aldrich; 0.5 mg/mL), fetuin (Sigma-Aldrich; 0.5 mg/mL), epidermal growth factor (Life Technologies; 10 ng/mL), dexamethasone (Sigma-Aldrich; 0.39 μg/mL), insulin (Sigma-Aldrich; 0.04 mg/mL), creatine (Sigma-Aldrich; 1 mM), pyruvate (Sigma-Aldrich; 100 μg/mL), uridine (Sigma-Aldrich; 50 μg/mL), and gentamycin (Life Technologies; 5 μg/mL) for 5 to 7 days.
Human myoblasts, defined as CD56 + CD146 + CD82 + CD45 -CD34 -, were sorted by fluorescence-activated cell sorting (FACS) using an Aria cytometer (BD Biosciences).
Human monocytes were obtained from peripheral blood of healthy donors after Ficoll-Paque gradient separation (GE Healthcare Bio-Sciences AB, Uppsala, Sweden) and positive selection by FACS using a mouse anti-human CD14 antibody (BD). The purity of human monocytes was more than 95% and their viability more than 99% using the trypan blue exclusion test.
Human myoblasts, defined as CD56 + CD146 + CD82 + CD45 -CD34 -, were sorted by fluorescence-activated cell sorting (FACS) using an Aria cytometer (BD Biosciences).
Human monocytes were obtained from peripheral blood of healthy donors after Ficoll-Paque gradient separation (GE Healthcare Bio-Sciences AB, Uppsala, Sweden) and positive selection by FACS using a mouse anti-human CD14 antibody (BD). The purity of human monocytes was more than 95% and their viability more than 99% using the trypan blue exclusion test.
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