Flow cytometric analysis for cell surface markers were performed by blocking expanded cells first with FCR Blocking Reagent (1:5; Miltenyi Biotec Inc) and followed by incubating for 20 min at 4 °C with the following antibodies: anti-CD34-PE, and anti-CD133/2 FITC (all from Miltenyi Biotec Inc), CXCR4, CD31, CD14, CD161, MHC Class-I, MHC Class-II, LFA-1, VCAM-1, CD45, CD69, CD18, CD36, CD3, CXCR2, CD71, CD45R and Isotype controls (IgG1 and IgG2a) were purchased from BD Biosciences, (USA). After incubation, cells were washed with MACS sorting buffer and analyzed using a FACS Calibur flowcytometer (Becton Dickinson, Heidelberg, Germany). Dead cells were excluded via propidium iodide staining. At least 20,000 events were acquired. Data analysis was performed with BD Cell Quest software.
Mhc class 1
Manufactured by BD
Sourced in United States
MHC Class I is a type of cell surface protein that plays a crucial role in the immune system. It is responsible for presenting peptide fragments derived from intracellular proteins to cytotoxic T cells, enabling the recognition and elimination of infected or abnormal cells. The MHC Class I molecule is composed of a heavy chain and a light chain (beta-2 microglobulin), and its function is essential for the proper functioning of the adaptive immune response.
Automatically generated - may contain errors
Lab products found in correlation
Facscalibur, by BD (3 mentions)
Mhc class 2, by BD (2 mentions)
Cellquest software, by BD (2 mentions)
Anti cd34 pe, by Miltenyi Biotec (1 mentions)
Cxcr4, by Miltenyi Biotec (1 mentions)
Lfa 1, by BD (1 mentions)
Vcam 1, by BD (1 mentions)
Cxcr2, by BD (1 mentions)
Facscalibur flow cytometer, by BD (1 mentions)
Fcr blocking reagent, by Miltenyi Biotec (1 mentions)
Hla dr, by R&D Systems (1 mentions)
Facscalibur system, by BD (1 mentions)
Cd105, by BD (1 mentions)
Cd106, by BD (1 mentions)
5 protocols using mhc class 1
1
Comprehensive Immunophenotyping of Cell Populations
2
Characterizing Cell Surface Markers
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Established clonal cells at passage 4–9 were subjected to flow cytometry for analysis of cell surface marker proteins. Briefly, the cells were washed twice with PBS, harvested by treatment with trypsin/EDTA, then incubated with the fluorescein isothiocyanate (FITC) or phycoerythrin (PE)-conjugated antibodies. Cells were then analyzed using a FACSCalibur system (BD Biosciences, San Jose, CA, USA), after which data were analyzed using the Cell Quest software (BD Biosciences). The antibodies used for flow cytometric analysis were as follows: CD29 (BD Biosciences), CD44 (BD Biosciences), CD73 (BD Biosciences), CD90 (R&D Systems, Minneapolis, MN, USA), CD105 (BD Biosciences), and MHC-class I (BD Biosciences) for mesenchymal markers, CD14 (BD Biosciences), CD34 (BD Biosciences), CD45 (BD Biosciences), CD117 (BD Biosciences), and HLA-DR (R&D Systems) for hematopoietic markers, AQP5 (Bioss, Woburn, MA) and CD24 (BD Biosciences) for epithelial markers, and SOX2 (Cell Signaling Technology, Danvers, MA) and OCT4 (R&D Systems) for embryonic markers. Isotype-matched control antibodies were used in each antibody analysis.
3
Immunophenotyping of BM-MSCs
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The BM-MSCs were harvested on the third passage and underwent flow cytometric analysis with the FACS Calibur device (BD Biosciences, San Jose, CA). In this immunophenotyping, antibodies against CD29, CD45, CD54, CD90, CD106, and MHC Class I (BD Biosciences) were evaluated (Figure 1 ).
4
Immunophenotyping of rBM-MSCs by Flow Cytometry
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Undifferentiated rBM-MSCs were subjected to flow cytometry analysis (FACS Calibur [BD Biosciences, San Jose, CA]). The cell suspension was spun at 1000 RPM for 5 min and the supernatant was decanted. The pellet was resuspended in 1X phosphate buffered saline (PBS). The cells were counted with a haemocytometer. The desired total number of cells was added to a flow tube (0.5-1 × 10 6 per sample). The cells were washed by adding ~1 mL 1X PBS to the flow tube. The cell suspension was spun at 1000 RPM for 5 min and the supernatant was decanted. The tube was gently tapped to loosen the cell pellet. An appropriate amount of staining buffer (50 µL per 1 × 10 6 cells) was added and 1 × 10 6 cells (50 µL) were added to the desired number of flow tubes. Finally, immunophenotyping analysis was performed for the antigens CD29, CD45, CD54, CD90, CD106, MHC class-I and MHC class-II (BD Biosciences).
5
BM-MSCs Immunophenotyping Analysis
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The BM-MSCs were harvested on the third passage and underwent flow cytometric analysis with the FACS Calibur device (BD Biosciences, San Jose, CA). In this immunophenotyping, antibodies against CD29, CD45, CD54, CD90, CD106, and MHC Class I (BD Biosciences) were evaluated (Figure 1).
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