Blood, spleen, and BM cells from hu-mice were stained with antibodies and analyzed using a BD Accuri C6 Plus (BD Biosciences). The antibodies used to detect human cells included APC-conjugated anti-human CD45 (555485, BD PharmingenTM), FITC-conjugated anti-mouse CD45 (553080, BD PharmingenTM), APC-conjugated anti-human CD4 (555349, BD PharmingenTM), PE-conjugated anti-human CD8 (555367, BD PharmingenTM), PE-conjugated anti-human CD19 (302208, BioLegend), and PE-conjugated anti-human CD33 (303403, BioLegend). For immunocy-tochemistry, anti-CD45 (ab8216, Abcam) and anti-CD34 (MAB72271, R&D Systems) antibodies were used, and proper isotype-matched IgG antibodies were used to detect the primary signals. Flow cytometric data were analyzed using the CSamplerTM Plus software program (BD Bio-sciences).
Apc conjugated anti human cd45
Manufactured by BD
Sourced in United States
The APC-conjugated anti-human CD45 is a laboratory reagent used to identify and quantify CD45-positive cells in flow cytometry applications. CD45 is a transmembrane glycoprotein expressed on the surface of most hematopoietic cells. The APC (Allophycocyanin) fluorescent dye is conjugated to the anti-CD45 antibody, allowing for the detection and enumeration of CD45-expressing cells.
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Lab products found in correlation
Ab8216, by Abcam (1 mentions)
Pe conjugated anti human cd8, by BD (1 mentions)
Pe conjugated anti human cd33, by BioLegend (1 mentions)
Fitc conjugated anti mouse cd45, by BD (1 mentions)
Pe conjugated anti human cd19, by BioLegend (1 mentions)
Accuri c6 plus, by BD (1 mentions)
Fitc conjugated anti human cd34, by BD (1 mentions)
Facsdiva software, by BD (1 mentions)
Lsr 2, by BD (1 mentions)
Pe cy7 conjugated anti human cd34, by BD (1 mentions)
Pe cy7 conjugated anti human cd19, by BD (1 mentions)
Pe conjugated anti human cd 11b, by BD (1 mentions)
Facscanto 2, by BD (1 mentions)
Facsaria 2, by BD (1 mentions)
Apc conjugated anti human cd3, by BD (1 mentions)
Collagenase type 1, by Merck Group (1 mentions)
Retronectin t100a, by Takara Bio (1 mentions)
Percp cy5.5 conjugated anti human cd3, by BD (1 mentions)
Tnf α and ifn γ, by Thermo Fisher Scientific (1 mentions)
Hyaluronidase, by Merck Group (1 mentions)
4 protocols using apc conjugated anti human cd45
1
Immunophenotyping of Humanized Mice
2
Quantification of Endothelial Progenitor Cells
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The number of EPCs was tested by fluorescence-activated cell sorting (BD LSR II with FACS Diva Software, BD Biosciences, USA). Two hundred microliters of whole blood was transferred to polystyrene tubes for testing with the following antibodies added to the samples: APC-conjugated anti-human CD45, FITC-conjugated anti-human CD34 (BD Biosciences, USA), and PE-conjugated anti-human KDR/VEGF-2 (BD Biosciences, USA). The experimental protocol can be found elsewhere (13 (link)).
3
Flow Cytometric Analysis of Human Cells
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Human cells in the BM and PB of recipient mice were stained with Lineage mixture (biotinylated anti-human CD14, CD3, CD56, CD19 and CD235a), PE-Cy7-conjugated anti-human CD34, APC-conjugated anti-human CD45, PE-conjugated anti-human CD38, biotinylated anti-human CD7, biotinylated anti-human CD10, PE-conjugated anti-human CD62L, APC-Cy7-conjugated anti-human CD3, PE-conjugated anti-human CD11b, or PE-Cy7-conjugated anti-human CD19 (BD Biosciences, San Jose, CA, USA). They were then analyzed on a FACS Canto II or sorted on a FACS Aria II (BD Biosciences).
4
Cell Culture and Antibody Conditions
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HCT116, NCI-H630 and RKO cells were cultured in RPMI 1640 medium supplemented with 10 % FBS, 2 mM Ultraglutamine and 100U/ml penicillin/streptomycin (Lonza, BioWhittaker). The packaging cell line Phoenix-A and HEK 293 T cells were cultured in DMEM medium (Lonza, BioWhittaker) supplemented with 10 % FBS, 2 mM Ultraglutamine and 100U/ml penicillin/streptomycin. TNF-α and IFN-γ (Peprotech) were used for in vitro stimulation of cell lines.
Monoclonal antibodies used in this study were as follows: anti-CD3 mAB OKT3 (EB16-0037-85, eBioscience), PE-conjugated anti-human CX3CR1 (D070-5, MBL), allophycocyanin (APC) conjugated anti-human CD3 (555342, BD Biosciences), APC-Cy7 conjugated mouse anti-human CD8 (557852, BD Biosciences), PerCP-Cy5.5-conjugated anti-human CD3 (560835, BD Biosciences), APC conjugated anti-human CD45 (560973, BD Biosciences), PE-Cy7 conjugated anti-human CD4 (557852, BD Biosciences). Other reagents included RetroNectin (T100-A, Takara); recombinant human IL-2 (202-IL, R&D Biosystems); Collagenase Type I (C0130, Sigma); DNAse I (D5025, Sigma), Hyaluronidase (H3506, Sigma).
Monoclonal antibodies used in this study were as follows: anti-CD3 mAB OKT3 (EB16-0037-85, eBioscience), PE-conjugated anti-human CX3CR1 (D070-5, MBL), allophycocyanin (APC) conjugated anti-human CD3 (555342, BD Biosciences), APC-Cy7 conjugated mouse anti-human CD8 (557852, BD Biosciences), PerCP-Cy5.5-conjugated anti-human CD3 (560835, BD Biosciences), APC conjugated anti-human CD45 (560973, BD Biosciences), PE-Cy7 conjugated anti-human CD4 (557852, BD Biosciences). Other reagents included RetroNectin (T100-A, Takara); recombinant human IL-2 (202-IL, R&D Biosystems); Collagenase Type I (C0130, Sigma); DNAse I (D5025, Sigma), Hyaluronidase (H3506, Sigma).
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