Antibodies (Abs) used for flow cytometry, PE mouse antihuman CD33 (Cat#: 555,450) and APC mouse antihuman CD45 (Cat#: 555,485) were purchased from BD Biosciences, PerCP/Cy5.5 antihuman CD19 (Cat#: 302,229) was purchased from Biolegend. Cytarabine (Cat#: BD8499), imatinib (Cat#: BD42606), and ibrutinib (Cat#:BD254580) were purchased from Bide Pharmatech (Shanghai, China). Epirubicin (Cat#: 56,390-09-1) was purchased from Shandong New Time Pharmaceutical Co., Ltd., Company (Shandong, China). Vincristine (Cat#: MB1298) was purchased from Melone Pharma (Dalian, China). Cladribine (Cat#:CSN10004) was purchased from CSNpharma (Shanghai, China).
Apc mouse anti human cd45
Manufactured by BD
Sourced in Germany
The APC mouse anti-human CD45 is a lab equipment product that functions as a cell surface marker. It is designed to detect the expression of the CD45 protein on human cells.
Automatically generated - may contain errors
Lab products found in correlation
Fitc rat anti mouse cd45, by BD (3 mentions)
Pe mouse anti human cd105, by BD (2 mentions)
Pe mouse anti human cd73, by BD (2 mentions)
Pe mouse anti human cd34, by BD (2 mentions)
Ibrutinib, by Bide Pharmatech (1 mentions)
Pe mouse anti human cd33, by BD (1 mentions)
Lsr 2, by BD (1 mentions)
Apc mouse anti human cd34, by BD (1 mentions)
Apc mouse anti human cd43, by BD (1 mentions)
Pe mouse anti human cd31, by BD (1 mentions)
Fitc conjugated mouse igg2a, by BD (1 mentions)
Pe conjugated mouse igg1κ, by BD (1 mentions)
Fitc mouse anti human cd34, by BD (1 mentions)
Cytoflex v2 b4 r2 flow cytometer, by Beckman Coulter (1 mentions)
Calibur flow cytometer, by BD (1 mentions)
Apc conjugated mouse igg1, by BD (1 mentions)
Accuri c6 cytometer, by BD (1 mentions)
Mitosox, by Thermo Fisher Scientific (1 mentions)
Lsrii ow cytometer, by BD (1 mentions)
8 protocols using apc mouse anti human cd45
1
Antibody Selection for Flow Cytometry
2
Phenotypic Characterization of Human Adipose-Derived Stem Cells
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Cells were harvested, washed and centrifuged for 5 min at 300 g. Pellets were resuspended in sterile complete media, and cell counts were determined. Approximately 500,000 cells were used for each reaction, including different cell types for comparison purposes (fibroblasts and hASCs). Cells were recorded on a BD LSR II flow cytometer (BD Biosciences, Oxford, UK) using BD FACSDiva software, and data were analyzed using FlowJo software (TreeStar., Ashland, OR, USA) [19 (link)]. The following panel of antibodes were used for hASC characterization: APC mouse anti-human CD45 (BD Biosciences catalog no #555485), PE mouse anti-human CD34 (BD Horizon, BD Biosciences, catalog no #562577), PE mouse anti-human CD73 (BD Biosciences catalog no #550257), PE mouse anti-human CD90 (Biosciences catalog no #555596) and PE mouse anti-human CD-105 (BD Biosciences catalog no #560839). The percentage of fluorescence in hASCs was determined.
3
Xenograft Modeling of Leukemia in NSG Mice
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NSG (NOD/Shi-scid/IL-2Rγnull) mice (in-house breeding; female, 4–6 weeks) were sublethally irradiated (250 rads), and each mouse was injected with 0.6 × 106 cells via lateral tail vein. Engraftment was assessed by flow cytometric measurement of human and mouse CD45 expression on the cell surface (APC mouse anti-human CD45 and FITC rat anti-mouse CD45, BD Biosciences). 15 mice were transplanted with MV4;11-C cells, 7 of which were assessed for engraftment. 10 mice were transplanted with MV4;11-KD cells, 5 of which were assessed for engraftment. For REH-C and REH-KD cells, 5 mice were transplanted with each cell line, and all were assessed for engraftment. The sample sizes were chosen based on efficiency of resource use, past experience with similar experiments, and pilot experiments to determine the effect size. No animal was excluded from the analysis, randomization was not used, and the investigator was not blinded. All animal procedures were conducted in accordance with the Guide for the Care and Use of Laboratory Animals (National Institute of Health, Bethesda, MD, USA) and were approved by the Institutional Animal Care and Use Committee at Johns Hopkins University.
4
Immunophenotyping of Differentiated Cells
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The differentiated cells in T12.5 flask were dissociated into single cells using 0.05% trypsin supplemented with 0.1% EDTA. The cells were washed, resuspended in a FACS washing buffer (PBS with 5% BSA and 2.5 mM EDTA) and then stained with the desired antibodies. The antibodies used in our study were: PE Mouse Anti-Human CD31 (1:25, BD), FITC Mouse Anti-Human CD34 (1:25, BD), APC Mouse Anti-Human CD34 (1:25, BD), APC Mouse Anti-Human CD43 (1:25, BD), APC Mouse Anti-Human CD45 (1:25, BD), FITC-conjugated mouse IgG2a (1:25, BD), APC-conjugated mouse IgG1 (1:25, BD) and PE-conjugated mouse IgG1κ (1:25, BD) were used as isotype-matched negative controls. Flow cytometry was performed on Calibur flow cytometer (BD) or CytoFLEX V2-B4-R2 Flow Cytometer (BECKMAN COULTER), and the data was analyzed using FlowJo software, version 10.0.7.
5
Xenograft Modeling of Leukemia in NSG Mice
6
Mitochondrial ROS in Human PBMCs
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After erythrocyte lysis with RBC Lysis Solution (MACS, Cat. number 130-094-183, Milteny Biotech, Germany), 200 μl of human PBMCs were labeled with 5 μL of APC-CD45 antibody (BD pharmingen APC-mouse anti human CD45. CAT number 555485 BD biosciences, NJ, USA) and 1 μM MitoSox (Thermo-Fisher Scientific, MA, USA). Fluorescence was measured in a C6 Accuri cytometer (BD Biosciences, NJ, USA) with a blue laser (488 nm) and FL3 filter (585/40 nm) (MitoSox Ex/Em = 510/580). 10,000 cells were analysed in each experiment, and the fluorescence registered was relativized to that of an internal control consisting of U937 cells that had undergone the same protocol.
7
Mouse Leukocyte Isolation and Staining
8
Characterization of Human Adipose-Derived Stem Cells
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Cells were harvested, washed, and centrifuged for 5 min at 300 g. Pellets were resuspended in sterile complete media and cell counts were determined. Approximately 500.000 cells were used for each reaction, including different cell types for comparison purposes ( broblasts and hASC). Cells were recorded on the BD LSR II ow cytometer (BD Biosciences, Oxford, UK) using BD FACSDiva software, and data were analyzed using FlowJo software (TreeStar., Ashland, OR, USA) (18). The following antibody panel was used for hASC characterization: APC mouse anti-human CD45 (BD Biosciences catalog nº #555485), PE mouse anti-human CD34 (BD Horizon, BD Biosciences, catalog nº #562577), PE mouse anti-human CD73 (BD Biosciences catalog nº #550257), PE mouse anti-human CD90 (Biosciences catalog nº #555596) and PE mouse anti-human CD-105 (BD Biosciences catalog nº #560839). The percentage of uorescence in hASC was determined in all cases.
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