Fitc conjugated cd45

Manufactured by BD

Sourced in Australia, United States

FITC-conjugated CD45 is a laboratory reagent used for the identification and enumeration of hematopoietic cells. CD45 is a pan-leukocyte marker expressed on the surface of all cells of hematopoietic origin. The FITC conjugation allows for the detection of CD45-positive cells using flow cytometry.

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Lab products found in correlation

Attune nxt, by Thermo Fisher Scientific (2 mentions) Pe conjugated cd105, by BD (1 mentions) 7 aad, by Beckman Coulter (1 mentions) Fitc conjugated f4 80, by BioLegend (1 mentions) A1 confocal imaging system, by Nikon (1 mentions) Slowfade diamond, by Thermo Fisher Scientific (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions) F4 80, by BD (1 mentions) Pe conjugated cd144, by BD (1 mentions) Apc conjugated cd90, by BioLegend (1 mentions) Facsaria 3, by BD (1 mentions) Accuri c6, by BD (1 mentions) Pe conjugated cd34, by BD (1 mentions) Pe conjugated cd73, by BD (1 mentions) Af 700 conjugated cd3, by BD (1 mentions) Apc conjugated cd105, by BioLegend (1 mentions) Facsdiva software, by BD (1 mentions) Lsrii flow cytometer, by BD (1 mentions) Apc conjugated cd44, by BD (1 mentions) Pe conjugated cd14, by BD (1 mentions)

4 protocols using fitc conjugated cd45

Adhesion Molecule Expression in hEC-U937 Co-culture

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hEC or HUVEC were stained with PE-conjugated E-selectin, PE-conjugated ICAM-1, and APC-conjugated VCAM-1 (all from BD Biosciences, Franklin Lakes, NJ).
U937 cells were suspended at a final concentration of 1×10⁶ cells/mL in media and plated on hEC layers pre-treated with 50 μg/mL histone for 1 h. The co-cultured cells were treated with cytosine D–arabinofuranoside (Ara-C; Hospira Pty Ltd., Mulgrave, Australia) for 24 h or without Ara-C for 48 h. Adherent and non-adherent U937 cells were collected separately and stained with FITC-conjugated CD45 (BD Biosciences), PE-conjugated CD105 (BD Biosciences), 7-AAD (Beckman Coulter, Fullerton, CA).

Yoo H.J., Lee J.S., Kim J.E., Gu J., Koh Y., Kim I, & Kim H.K. (2016). Extracellular Histone Released from Leukemic Cells Increases Their Adhesion to Endothelium and Protects them from Spontaneous and Chemotherapy-Induced Leukemic Cell Death. PLoS ONE, 11(10), e0163982.

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Immunofluorescent Analysis of Tumor Vasculature and Immune Cells

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Frozen 4T1 tumors were fixed in 4% paraformaldehyde followed by blocking and staining with an antibody to CD31 (BD Biosciences) overnight at 4ºC, as previously described51 (link). After overnight incubation, sections were washed and an Alexa Fluor 488 secondary antibody was added for 1 h at room temperature. For CD45 and F4/80 staining, tissue sections were prepared as described above and stained with FITC-conjugated CD45 (BD Pharmingen) and FITC-conjugated F4/80 (Biolegend) overnight at 4ºC. Sections were then counterstained with 4',6-diamidino-2-phenylindole (ThermoFisher Scientific), mounted using SlowFade Diamond (ThermoFisher Scientific), and cover-slipped. Tumor sections were imaged using a Nikon A1 Confocal Imaging System housed within the Advanced Cellular and Tissue Microscopy Core at HMRI.

Martinez J.O., Molinaro R., Hartman K.A., Boada C., Sukhovershin R., De Rosa E., Kirui D., Zhang S., Evangelopoulos M., Carter A.M., Bibb J.A., Cooke J.P, & Tasciotti E. (2018). Biomimetic nanoparticles with enhanced affinity towards activated endothelium as versatile tools for theranostic drug delivery. Theranostics, 8(4), 1131-1145.

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Cell Isolation and Flow Cytometry

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At desired time points, nonadherent and adherent cells were harvested separately by aspiration and dissociation with 0.5 mM EDTA for 15 min, respectively. The cells were then filtered through 40 μm strainers, pelleted, and stained with antibodies in PBE buffer (PBS + 0.5% BSA + 2 mM EDTA). Antibodies of flow cytometry were used as follows: PerCP-conjugated CD34 (BioLegend), FITC-conjugated CD45 (BD Biosciences), and APC-conjugated CD90 (BioLegend), PE-conjugated CD144 (BD Biosciences), and APC-conjugated CD73 (BD Biosciences). Detailed information on these antibodies is provided in Additional file 1: Table S1. All samples were analyzed using BD Accuri™ C6 (BD Biosciences) and Attune™ Nxt (ThermoFisher Scientific). Cell sorting was performed using a FACSAria™ III (BD Biosciences). The data of flow cytometry was analyzed using Flowjo software.

Thongsa-ad U., Wongpan A., Wongkummool W., Chaiwijit P., Uppakara K., Chaiyakitpattana G., Singpant P., Tong-ngam P., Chukhan A., Pabuprappap W., Wongniam S., Suksamrarn A., Hongeng S., Anurathapan U., Kulkeaw K., Tubsuwan A, & Bhukhai K. (2024). Improving hematopoietic differentiation from human induced pluripotent stem cells by the modulation of Hippo signaling with a diarylheptanoid derivative. Stem Cell Research & Therapy, 15, 60.

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Multiparameter Flow Cytometry Profiling

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Cultured cells (passage 1) were trypsinized and stained with a panel of antibodies for fluorescence-activated cell sorting (FACS) analysis. Approximately, 1 × 10⁵ cells were re-suspended in phosphate buffered saline (PBS) and incubated with IgG block for 5 minutes to block non-specific binding. The following antibodies were used: AF-700 conjugated CD3 (BD BioSciences, USA), PE conjugated CD14 (BD, Immunocytometry, USA), APC conjugated CD19 (BD BioSciences, USA), PE conjugated CD34 (BD, BioSciences, USA), APC conjugated CD44 (BD, Pharmingen, USA), FITC conjugated CD45 (BD Pharmingen, USA), PE conjugated CD73 (BD Pharmingen, USA), AF-700 conjugated CD90 (Biolegend, USA) and APC conjugated CD105 (Biolegend, USA). Cells were stained for 30 minutes at 4°C with the antibodies. After washing, samples were analyzed on a LSR II flow cytometer (BD, USA) and at least 10,000 events were acquired for each population. Data acquisition and analysis were performed using FACS DIVA software (BD Biosciences, USA). Unstained cells were used to establish flow cytometer settings. Debris and cells/particles with auto-fluorescence were removed by using a threshold on the forward scatter.

Choudhery M.S., Badowski M., Muise A., Pierce J, & Harris D.T. (2014). Donor age negatively impacts adipose tissue-derived mesenchymal stem cell expansion and differentiation. Journal of Translational Medicine, 12, 8.

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