Pe conjugated cd 133

Manufactured by Thermo Fisher Scientific

PE-conjugated CD-133 is a cell surface marker that is expressed on hematopoietic stem and progenitor cells. It can be used for the identification and isolation of these cell populations through flow cytometry or cell sorting techniques.

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Lab products found in correlation

Hyaluronidase type 4 s, by Merck Group (1 mentions) Apc conjugated cd31, by Thermo Fisher Scientific (1 mentions) Dnase 1 grade 2, by Roche (1 mentions) Liberase tm research grade, by Roche (1 mentions) 70 μm nylon cell strainer, by BD (1 mentions) Dispase, by Roche (1 mentions) Fluorescence activated cell sorter, by BD (1 mentions) Ficoll, by Merck Group (1 mentions) Fitc conjugated cd34, by Thermo Fisher Scientific (1 mentions) Egm 2, by Lonza (1 mentions)

3 protocols using pe conjugated cd 133

Flow Cytometric Analysis of Angiogenic Cell Types

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Subcutaneous matrigel plugs were digested for 60–90 minutes at 37 °C in an enzymatic cocktail composed of 25 μg/ml DNase-I, grade II (Roche, cat no. 10-104-159-001), 3 U/ml Dispase (Roche, cat no. 10-269-638-001), 3 U/ml Liberase TM Research grade (Roche, cat no. 05-401-119-001) and 25 μg/ml Hyaluronidase Type IV-S (Sigma, cat no. H3884). The isolated cells were filtered through a 70 μm Nylon cell strainer (BD Falcon, Bedford, MA) into a sterile 50 ml centrifuge tube and cells washed 3X with FACS Buffer (1X PBS without Ca/Mg, 2 mM EDTA and 0.5% BSA).
Isolated cells were analyzed by flow cytometry to determine their cellular characteristics. FITC-conjugated F4/80 (eBioscience, cat no.11-4801) and APC-conjugated CD31 (eBioscience, cat no. 17-0311) were used to distinguish endothelial cells from macrophages with additional labeling using eFluor450-conjugated CD34 (eBioscience, cat no. 48-0341), PE-conjugated CD-133 (eBioscience, cat no. 12-1331) or PERC-P conjugated CD45 (eBioscience, cat no. 45-0451). Antibodies were incubated with isolated cells in FACS buffer for 30 minutes at 4 °C in darkness. Cells were washed 3X with FACS buffer and fixed in 1% PFA in PBS for 10 minutes at room temperature in the absence of light. Fixed cells were washed in PBS and resuspended in 400 μl of FACS buffer for flow cytometry. Experiment performed three times.

Barnett F.H., Rosenfeld M., Wood M., Kiosses W.B., Usui Y., Marchetti V., Aguilar E, & Friedlander M. (2016). Macrophages form functional vascular mimicry channels in vivo. Scientific Reports, 6, 36659.

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Isolation of CD133+ Neural Progenitor Cells

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Three-week-old mice were anesthetized and euthanized in accordance with Tongji University institutional guidelines. The brains were quickly removed from the skull and put into cold PBS. After several washes with cold PBS at 37°C for 30 min, the tissue was dissected under a dissection microscope, and the walls of the lateral ventricles were obtained and enzymatically digested using Papain (Worthington LS003127) at 37°Cfor 30 min. Dissociated cells were labeled with phycoerythrin (PE)-conjugated CD133 (eBiosciences 12-1331) for 1 hr. After several PBS washes, labeled cells were sorted using the Becton Dickinson Fluorescence-Activated Cell Sorter to isolate CD133⁺ or CD133⁻ cells, followed by manual picking of single cells under the microscope.

Luo Y., Coskun V., Liang A., Yu J., Cheng L., Ge W., Shi Z., Zhang K., Li C., Cui Y., Lin H., Luo D., Wang J., Lin C., Dai Z., Zhu H., Zhang J., Liu J., Liu H., deVellis J., Horvath S., Sun Y.E, & Li S. (2015). Single-Cell Transcriptome Analyses Reveal Signals to Activate Dormant Neural Stem Cells. Cell, 161(5), 1175-1186.

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Isolation and Characterization of Endothelial Progenitor Cells

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Human blood samples were collected in heparin anticoagulant tubes through venipuncture, and peripheral blood mononuclear cells (PBMC) were obtained by density gradient centrifugation with Ficoll (1.077 g/ml; Sigma). At last, PBMC were incubated for 30 min at room temperature in a dark room with appropriate concentrations of the following monoclonal antibodies: Alexa Fluor 647 conjugated KDR (BD Biosciences), FITC conjugated CD34 (eBioscience), PE conjugated CD133 (eBioscience). Appropriate isotype controls were used. Samples were assessed by ow cytometer with FACS Cell Quest analysis software (Becton-Dickinson, CA, USA). At least 1,000,000 events were recorded in the mononuclear cell gate set on the SSC/FSC morphological plot, and the results were analyzed by FlowJo V10 software (Tree Star Inc, Ashland, USA).
Endothelial progenitor cell culture PBMC obtained by density gradient centrifugation were seeded on bronectin (Corning)-coated plates in order to evaluate cell's function. 2 million cells per well were seeded in 24-well plates, and 4 million cells per well were seeded in 12-well plates. Cells were cultured in EGM-2(Lonza), after 4 days, non-adherent cells were removed, and fresh mediums were added. The culture was maintained through day 21, and adherent cells were subjected to further examinations.

Zhu Y., Na-na Y., Zhu J., Fu C., Tian X., Zhang F., Zhao L., Zhu J., Lv K., & Wang C. (2020). The number of circulating endothelial progenitor cells from septic patients are associated with different infectious organisms.

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