Freshly prepared PBMCs were used. Subpopulations of T cells, B cells, natural killer (NK) cells and antigen-presenting cells (APC) were characterized by surface staining with fluorescence labelled anti-CD3, anti-CD4, anti-CD5, anti-CD8, anti-CD14, anti-CD16, anti-CD19, anti-CD25, anti-CD27, anti-CD38, anti-CD45RA, anti-CD45RO, anti-CD56 and anti-HLA-DR (BD Bioscience); anti-BDCA1, anti-BDCA2, anti-BDCA3, anti-BDCA4 and anti-slan (Miltenyi Biotec). Negative controls included directly labeled or unlabeled isotype-matched irrelevant antibodies (BD Biosciences). Freshly prepared CSF cells were used directly for FACS analysis. Fluorescence-labeled antibodies for surface staining were used as follows: anti-CD3, anti-CD4, anti-CD8, anti-CD14, anti-CD16, anti-CD19, anti-CD25, anti-CD27, anti-CD45RA, anti-CD45RO, anti-CD56 and anti-HLA-DR (BD Bioscience); anti-BDCA1 and anti-slan (Miltenyi Biotec). All cells were measured on a LSR-Fortessa (BD Biosciences) and evaluated by FACS-Diva Software (BD Bioscience).
Anti hla dr
Manufactured by BD
Sourced in United States
Anti-HLA-DR is a laboratory test used to detect the presence of HLA-DR antigen on the surface of cells. HLA-DR is a major histocompatibility complex (MHC) class II cell surface receptor that plays a crucial role in the immune response. The anti-HLA-DR assay is commonly used in research and clinical settings to analyze cell populations and their immune functions.
Automatically generated - may contain errors
Lab products found in correlation
Anti cd45, by BD (13 mentions)
Anti cd8, by BD (11 mentions)
Anti cd4, by BD (11 mentions)
Anti cd86, by BD (11 mentions)
Anti cd90, by BD (10 mentions)
Anti cd73, by BD (10 mentions)
Anti cd34, by BD (9 mentions)
Anti cd3, by BD (8 mentions)
Anti cd105, by BD (7 mentions)
Anti cd80, by BD (6 mentions)
Facscalibur, by BD (5 mentions)
Anti cd31, by BD (5 mentions)
Anti cd117, by BD (5 mentions)
Anti cd69, by BD (5 mentions)
Facscalibur flow cytometer, by BD (5 mentions)
Anti cd45ra, by BD (4 mentions)
Anti cd56, by BD (4 mentions)
Anti cd19, by BD (4 mentions)
Anti cd45ro, by BD (4 mentions)
Facscanto, by BD (4 mentions)
55 protocols using anti hla dr
1
Comprehensive Immune Cell Profiling
2
Flow Cytometric Characterization of MSCs and HUVECs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
WJ-MSCs and HUVECs, respectively, at the eighth and the sixth passage, were treated with 0.05% trypsin–EDTA and collected; 106 cells per sample were incubated with 1 μg of the specific antibody, conjugated with fluorescein isothiocyanate (FITC), phycoerythrin (PE), allophycocyanin (APC), phycoerythrin-cyanine 5.5 (PE Cy5.5), or Alexa Fluor 488 for 30 min at 4°C in the dark. WJ-MSCs were stained using the following antibodies: anti-CD31, anti-CD73, anti-CD13, anti-CD90, anti-CD117, anti-CD14, anti-CD34, anti-CD105, anti-CD146, anti-CD133, anti-CD144, anti-ESA, anti-HLA-ABC, anti-HLA-DR, anti-CD45 (Becton Dickinson [BD], San Jose, CA), anti-CD29, anti-CD44, and anti-CD166 (Ancell, Bayport, MN). HUVECs were stained with anti-CD146 (BD) and anti-CD144 (Acris Antibodies, San Diego, CA). After incubation, cells were washed and acquired with a flow cytometer (FACS Calibur; BD), collecting 10,000 events per sample. Data were analyzed by the FlowJo software v8.8.6 (TreeStar, Ashland, OR). The mean fluorescence intensity (MFI) ratio values were calculated (i.e., dividing the MFI of positive events by the MFI of negative events).22 (link),23 (link)
3
Immunophenotypic Analysis of Peripheral Lymphocytes in COVID-19 Patients
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Peripheral blood white blood cells (WBC) of all participants were isolated from their hemogram tubes with EDTA taken for their routine tests by using erythrocyte lysing solution (155 mM NH4Cl; 10 mM KHCO3; 0,1 mM EDTA; pH:7.3). The following fluorochrome labeled monoclonal antibodies (mAb) and isotype-matched controls were used for two–three color phenotypic analysis: anti-IgG1, anti-IgG2a, anti-CD45, anti-CD3; anti-CD4; anti-CD8; anti-CD16; anti-CD56; anti-CD19; anti-HLA-DR; anti-CD69, (Becton&Dickinson Corp, San Jose, CA, USA). Cells were acquired and analyzed using CellQuest software on a FACS scan flow cytometer (Becton Dickinson Inc, San Jose, CA, USA). Lymphocytes were gated according to their forward and side scatter characteristics and their specific CD markers. Markers were evaluated as percentages. In 13 of 20 patients, peripheral lymphocyte subset analysis was repeated 1 week later. A total of 10 KTR without COVID-19 was used as control group.
4
Phenotypic Characterization of Treated Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Treated cells were characterized using the following monoclonal antibodies: anti-HLA-ABC (Invitrogen, Waltham, MA, USA), anti-PD-L1 (Becton Dickinson, Franklin Lakes, NJ, USA), and anti-HLA-DR (Becton Dickinson, Franklin Lakes, NJ, USA), conjugated with PE, Brilliant Violet 421. The staining was divided into two panels to avoid overlapping of emission spectra. Positive cells were distinguished from negative cells using unstained samples. FACS measurement was performed on the BD FACSCelesta Cell Analyzer.
5
Characterization of hPDLSCs by Flow Cytometry
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
hPDLSCs at the second passage were collected; 5 × 105 cells per sample were incubated with 1 μg of the specific antibody, conjugated with fluorescein isothiocyanate, phycoerythrin, allophycocyanin, phycoerythrin-cyanine 5.5, or Alexa Fluor 488 for 30 min at 4 °C in the dark. hPDLSCs were stained using the following antibodies: anti-CD13, anti-CD29, anti-CD44, anti-CD45, anti-CD105, anti-CD166 (Ancell, MN, USA), anti-CD14, anti-CD133 (BergischGladbach, Germany), anti-CD73, anti-CD90, anti-CD117, anti-CD146, anti-CD271, anti-Sox2, anti-HLA-DR, anti-SSEA4, anti-OCT3/4 (Becton Dickinson, BD, San Jose, CA, USA), anti-CD144 (Acris Antibodies, Herford, Germany) and anti-CD34 (Beckman Coulter, Fullerton, CA, USA). After incubation, cells were acquired with a flow cytometer (FACS Calibur; BD). Data were analyzed by the FlowJo software v8.8.6 (TreeStar, Ashland, OR, USA) [17 (link)].
6
Phenotypic Profiling of Immune Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Whole blood samples were collected in EDTA tubes by peripheral venipuncture. Peripheral blood mononuclear cells (PBMCs) were isolated using Ficoll-Paque (GE Health Care Life Sciences, Uppsala, Sweden), washed twice in PBS, and stained with commercial fluorochrome-conjugated anti-human monoclonal antibodies (mAbs): anti-CD4, anti-CD8, anti-CD20, anti-CD16, anti-CD25, anti-CD28, anti-CD38, anti-CD45RA, anti-CD45R0, anti-CD56, anti-CD69, anti-HLA-DR, and anti-programmed death receptor-1 (PD-1) (Becton-Dickinson, Holdrege, NE, USA) as previously described [24 (link)]. After incubation with the antibodies for 30 min at 4 °C, cells were washed and fixed in 1% paraformaldehyde (Sigma Chemical Co., St. Louis, MO, USA). Samples were acquired by FACSCanto flow cytometer and analyzed with the BD FACSDiva Software (BD Biosciences, San Jose, CA, USA).
7
Phenotypic Characterization of MenSCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The expression of isolated MenSCs surface markers was evaluated using fluorescence-activated cell sorting (FACS). Briefly, 5 × 105 cells were collected and washed twice with stain buffer (BD Biosciences, San Jose, CA, USA). MenSCs were incubated in the dark for 20 min with the following primary antibodies: PE-conjugated CD29, CD34, CD45, CD73, CD90, CD105, CD117, and HLA-DR (Becton Dickinson, Franklin Lakes, NJ, USA). The stained cells were washed twice with stain buffer, resuspended in 500 μl of stain buffer and then analyzed using a FC500 flow cytometer (Beckman Coulter, Brea, CA, USA). IgG1 (Becton Dickinson, Franklin Lakes, NJ, USA) was used as an isotype control for the anti-CD29, anti-CD34, anti-CD45, anti-CD73, anti-CD90, anti-CD105, and anti-CD117 antibodies. IgG2a (Becton Dickinson) was used as the isotype control for the anti-HLA-DR antibody. The results were analyzed using FlowJo software (Tree Star, Inc., Ashland, OR, USA).
8
Characterizing Macrophage Differentiation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Differentiation of MACs in response to infection (MOI 300) was assessed via flow cytometry. Detached cells were fixed in 1% PFA, blocked with 10% FCS and incubated with the following monoclonal murine human-specific Abs for surface antigens: anti-CD14, anti-CD45, anti-HLA-DR, anti-CD31 (Becton Dickinson) and anti-VEGF-R2 (R&D Systems). For detection of intracellular antigens, cells were permeabilized with Cytofix Cytoperm and incubated with anti-CD68 (Becton Dickinson). In each experiment, control groups were stained with immunoglobulin isotype control Abs [mouse IgG2aκ, IgG1κ, IgG2bκ, (Becton Dickinson) and mouse IgG1 (R&D Systems)]. For assessment of cell viability, MACs isolated from Matrigel structures were incubated with 7-amino-actinomycin D (7AAD) staining solution (eBiosciences) and fixated with 1% PFA. Unstained cells were used as negative controls. Cells were analysed on a fluorescence-activated cell sorter (FACS Canto, Becton Dickinson) and results were analysed using Flowing Software (Terho, 2012) .
9
Tetramer-Based Quantification of Antigen-Specific CTLs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Peptide MRP3 765 -specific tetramer was used for the detection of peptide Mizukoshi et al. Phase I Trial of MRP3-Derived peptide for HCC. vaccine-induced CTLs. Peptide HIVenv 584 -specific tetramer was used as a negative control of tetramer assay. All tetramers were purchased from Medical Biological Laboratories Co., Ltd. (Nagoya, Japan). PBMCs were stained with anti-CD8-APC (Becton Dickinson, Tokyo, Japan), anti-CCR7-FITC (eBioscience, Tokyo, Japan), anti-CD45RA-PerCP-Cy5.5 antibodies (eBioscience, Tokyo, Japan), and with tetramer-PE for 30 min at room temperature. Cells were washed, fixed with 0.5% paraformaldehyde/PBS, and analyzed using a Becton Dickinson FACSAria II system. In the tetramer assays with the negative control tetramer, we did not observe more than 0.03% tetramer-positive cells in any assay. Based on the results of the negative control, responses to MRP 765 -specific tetramer were considered positive if more than 0.03% tetramer positive cells were detected. At least 1,000,000 PBMCs were acquired for each tetramer assay. For the detection of Tregs and MDSCs, the following anti-human monoclonal antibodies were used: anti-CD4 (Becton Dickinson), anti-CD14 (Becton Dickinson), anti-CD25 (Becton Dickinson), anti-CD127 (Becton Dickinson), and anti-HLA-DR (Becton Dickinson).
10
Multiparametric Flow Cytometry Analysis
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!