Cultured cells were harvest, non specific binding was blocked and the cells were stained with different combinations of the following directly-conjugated antibodies: CD105 (cat no. 561443, BD Pharmingen), CD73 (cat no. 550257, BD Pharmingen), CD90 (cat no. 559869, BD Pharmingen), CD34 (cat no. 555821, BD Pharmingen), CD14 (cat no. 345785, BD), CD19 (cat no. 555415, BD Pharmingen), CD31 (cat no. 555445, BD Pharmingen), CD45 (cat no. 345808, BD), CD146 (cat no. 550315, BD Pharmingen), HLA-DR (cat no. 555558, BD Pharmingen) and HLA class I (cat no. 555553, BD Pharmingen) as described before5 (link). 7-amino-actinomycin D (Sigma Aldrich) was used for dead cell exclusion. Samples were analyzed on a FACSCalibur (BD Bioscience) and data acquisition and analysis were performed using CellQuest (BD Bioscience) and FlowJo software (Tree star, Ashland, Oregon, USA) respectively. Results are presented as percent positively stained cells.
Hla class 1
Manufactured by BD
Sourced in United States
The HLA class I is a lab equipment product. It is used to detect and analyze the presence of human leukocyte antigen (HLA) class I molecules. HLA class I molecules play a crucial role in the immune system by presenting antigens to cytotoxic T cells. The HLA class I equipment provides a tool for researchers and clinicians to study the expression and function of these important proteins.
Automatically generated - may contain errors
Lab products found in correlation
Hla dr, by BD (3 mentions)
Cd133, by BD (2 mentions)
7 aminoactinomycin d, by Merck Group (1 mentions)
Facscalibur, by BD (1 mentions)
Cellquest, by BD (1 mentions)
Cd105, by BD (1 mentions)
Cd146, by BD (1 mentions)
Cd106, by BD (1 mentions)
Cd105, by Bio-Rad (1 mentions)
Cd166, by Bio-Rad (1 mentions)
Cxcr4, by BD (1 mentions)
G46 2, by BD (1 mentions)
Facscanto 2, by BD (1 mentions)
Cd11a, by BD (1 mentions)
Cd44v6, by BD (1 mentions)
Anti cd10, by BD (1 mentions)
Her 2 neu, by BD (1 mentions)
Cd206, by BD (1 mentions)
Facscanto, by BD (1 mentions)
4 protocols using hla class 1
1
Phenotypic Characterization of Cell Cultures
2
Isolation and Characterization of Mesenchymal Stem Cells
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Bone marrow aspiration was approved by the Inha University Hospital Institutional Review Board (IRB) and written informed consent was obtained from a healthy donor (IRB number #10-51). Highly homogeneous clonal MSCs were isolated from BM as described previously (30 (link)). Markers of MSCs were determined by flow cytometry using a number of specific antibodies, including CD14 (BD Biosciences, San Diego, CA, USA), CD29 (Serotech, Kidlington, UK), CD31 (Serotec), CD34 (BD Biosciences), CD44 (Serotec), CD45 (BD Biosciences), CD49f (BD Biosciences), CD73 (BD Biosciences), CD90 (BD Biosciences), CD105 (Serotec), CD106 (BD Biosciences), CD133 (BD Biosciences), CD166(Serotec), HLA-DR (BD Biosciences), HLA-Class I (BD Biosciences), CXCR4 (BD Biosciences), and Oct-4 (Cell Signaling Technology, Danvers, MA, USA). MSCs were positive for CD29, CD44, CD49f, CD73, CD90, CD105, CD166, HLA-Class I, and Oct-4 but were negative for C14, CD31, CD34, CD45, CD106, CD133, CXCR4, and HLA-DR (data not shown). For the assessment of differentiation potential, adipogenic, osteogenic, and chondrogenic differentiations were induced as described (6 (link)). MSCs isolated were successfully differentiated into these 3 mesenchymal cell types, indicating the multilineage differentiation potential (data not shown).
3
Flow Cytometry Characterization of Malignant Plasma Cells
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Material from myeloma and plasma cell leukemia patients was obtained as a waste product from a diagnostic procedure that had been approved by the hospital’s medical ethical committee. Bone marrow cells were stained for CD38 (HB7, BD), CD138 (CLB-1D4, Sanquin), cytoplasmic kappa light chain (G20-193, BD), cytoplasmic lambda light chain (1–555–2, BD), HLA-class I (G46-2.6, BD) or HLA-E (3D12, eBioscience). HLA expression was determined on CD38high cells. All flow cytometry-based assays were performed on a BD FACS Canto II. Data were analyzed with BD FACSDiva v6.1.2 or FlowJo 7.6.
4
Characterization of Cell Surface Markers
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The following fluorescein (FITC)-, allophycocyanin (APC)-or phycoerythrin (PE)-conjugated mouse anti-human monoclonal antibodies (mAbs) were used: Anti-CD10, -CD11a, CD11c, -CD18, -CD33, -CD40, -CDD44std, -CD44v5, -CD44v6, -CD54, -CD61, -CD62P, -CD86, -CD133, -CD206 (MR), -EGFR, -Her-2/neu, -HLA-DR, HLA class I, -CCR5, -CCR6, Her-2/neu all from BD Pharmingen (San Diego, CA, USA); anti-CD29, -CD36, -CD51, -CD58 from Immunotech (Marseille, France); anti-c-MET, -CCR1, -CCR2, -CCR3, -CCR7, -CXCR1, -CXCR2, -CXCR4 from R&D (Abington, UK); anti-Tag72, -Mucin1 (EMA, CD227), -EMMPRIN from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA) and anti-Epithelial Antigen, -Epithelial Membrane Antigen (EMA) from DAKO (Heverlee, Belgium). Isotype controls included appropriate FITC-, APC- or PE-labeled mouse IgG1, IgG2a or IgG2b. Cells were incubated with mAbs or isotype controls for 20 min at 4°C, washed, resuspended in PBS and analyzed by flow cytometry (FACS Canto; BD Biosciences Immunocytometry Systems, San Jose, CA, USA) using FACS DiVa software.
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