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Pe anti human cd45

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PE anti-human CD45 is a fluorescent-labeled antibody that specifically binds to the CD45 antigen expressed on human leukocytes. CD45 is a protein tyrosine phosphatase that plays a critical role in the regulation of signal transduction in immune cells.

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Lab products found in correlation

Ma900 sorter, by Sony (2 mentions) Apc anti human cd117, by BioLegend (2 mentions) Ficoll, by BD (2 mentions) Antihuman cd42b apc, by BioLegend (2 mentions) Apc and percp cy5.5 labeled, by BioLegend (2 mentions) Apc anti human cd8, by BioLegend (2 mentions) Facscalibur, by BD (2 mentions) Fitc anti human cd34, by BioLegend (2 mentions) Pe anti human cd206, by BioLegend (2 mentions) Apc anti human cd86, by BioLegend (2 mentions) Pe anti human cd86, by BioLegend (2 mentions) Cellquest software, by BD (2 mentions) Staining buffer, by BD (2 mentions) Apc anti human cd206, by BioLegend (2 mentions) Alexa fluor 488 anti human cd11b, by BioLegend (2 mentions) Alexa fluor 488 anti human cd105, by BioLegend (2 mentions) Accutase, by Nacalai Tesque (2 mentions) Human trustain fcx fc receptor blocking solution, by BioLegend (2 mentions) Fitc anti human cd4, by BioLegend (2 mentions) Apc anti human cd56, by BioLegend (2 mentions)

Close spelling variants of this product

Anti-CD45 (228 citations) CD45-PE (78 citations) Anti-CD45-PE (33 citations) Anti-human CD45 (24 citations) PE-Cy7 anti-human CD45 (10 citations) PE-anti-human CD45 antibody (4 citations) PE anti-human CD45 (clone 2D1) (3 citations) PE)-conjugated anti-human CD45 antibody (3 citations) PE)/Cyanine 7 anti-human CD45 (3 citations) PE/Cy7 anti-human CD45 Antibody (2 citations)

16 protocols using pe anti human cd45

1

Isolation and RNA-Seq Analysis of Leukemia Cells

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Peripheral blood was taken under written informed consent from patients during routine blood draws at screening and different timepoints during the first cycle of treatment with SDNX-5613 within the AUGMENT-101 clinical trial (NCT04065399). These studies were conducted in accordance to the Declaration of Helsinki and were approved by an institutional review board at the Dana-Farber Cancer Institute (IRB: #01-206). Peripheral blood mononuclear cells (PBMCs) were subsequently isolated using Ficoll (BD Bioscience) gradient centrifugation, viably frozen, and banked at the Dana-Farber Cancer Institute, Boston, MA. For longitudinal analysis, samples were thawed, washed twice in PBS, and stained with anti-human CD45 (PE) (Biolegend, 304007) and anti-human CD117 (APC) (Biolegend, 313205). CD45-low/CD117+ leukemia cells were FACS sorted (Sony MA900 sorter) and subsequently processed for RNA-Seq (see methods section on RNA-Seq).
Soto-Feliciano Y.M., Sánchez-Rivera F.J., Perner F., Barrows D.W., Kastenhuber E.R., Ho Y.J., Carroll T., Xiong Y., Anand D., Soshnev A.A., Gates L., Beytagh M.C., Cheon D., Gu S., Liu X.S., Krivtsov A.V., Meneses M., de Stanchina E., Stone R.M., Armstrong S.A., Lowe S.W, & Allis C.D. (2022). A Molecular Switch between Mammalian MLL Complexes Dictates Response to Menin–MLL Inhibition. Cancer Discovery, 13(1), 146-169.
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2

Murine Monoclonal Antibody Labeling Protocol

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Murine monoclonal antibodies (all IgG1κ): antihuman CD42b-APC (BioLegend), antihuman CD45-PE (BioLegend), antihuman/mouse C3/C3b/iC3b-FITC and unlabeled (Cedarlane), antihuman CD11b-PerCP/Cy5.5 and -PE (BioLegend), antihuman CD62P-PE/Cy5 (BioLegend), and goat antimouse-AF488. Isotype controls: APC- and PerCP/Cy5.5-labeled (BioLegend), PE-labeled (BioLegend), and FITC-labeled (Cedarlane).
Blatt A.Z., Saggu G., Cortes C., Herbert A.P., Kavanagh D., Ricklin D., Lambris J.D, & Ferreira V.P. (2017). Factor H C-Terminal Domains Are Critical for Regulation of Platelet/Granulocyte Aggregate Formation. Frontiers in Immunology, 8, 1586.
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3

Immunophenotypic Analysis of Tumor Cells

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The following mAbs were used for phenotypic analysis: i. biotin-SP-conjugated AffiniPure rabbit anti-human IgG (H+L) and ii. biotin-SP-conjugated AffiniPure rabbit anti-mouse IgG (H+L) (Jackson, West Grove, PA); iii. biotin-conjugated recombinant human FOLR1 (RD Systems, Minneapolis, MN / Thermo Scientific, Rockford, IL); i.v. streptavidin-APC (BD, San Jose, CA); v. BD ViaProbe (7-AAD); vii. anti-human CD3-FITC, anti-human CD4-FITC, anti-human CD8-FITC (eBioscience). In adoptive immunotherapy experiments, T cells from peripheral blood were obtained via retro-orbital bleeding and stained for the presence of human CD45, CD3 and CD8 using anti-human CD45-PE, anti-human CD3-PerCP/Cy5.5 and anti-human CD8-APC (Biolegend, San Diego, CA). After gating on the human CD45+ population, the CD3+ and CD8+ subsets were quantified using TruCount tubes (BD Biosciences) with known numbers of fluorescent beads as described in the manufacturer's instructions. Tumor cell surface expression of FR was detected by Mov18/ZEL antibody (Enzo Life Sciences). Flow samples were run using BD FACS Canto and cytometric data analyzed by FlowJo 7.6.5 software.
Schutsky K., Song D.G., Lynn R., Smith J.B., Poussin M., Figini M., Zhao Y, & Powell D.J. (2015). Rigorous optimization and validation of potent RNA CAR T cell therapy for the treatment of common epithelial cancers expressing folate receptor. Oncotarget, 6(30), 28911-28928.
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4

CD45lo/CD117+ Leukemia Cell Isolation

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Peripheral blood was taken under written informed consent from patients during routine blood draws at screening and different timepoints during the first cycle of treatment with SDNX-5613 within the AUGMENT-101 clinical trial (NCT04065399). These studies were conducted in accordance with the Declaration of Helsinki and were approved by an institutional review board (IRB) at the Dana-Farber Cancer Institute (IRB: #01-206). Peripheral blood mononuclear cells were subsequently isolated using Ficoll (BD Biosciences) gradient centrifugation, viably frozen, and banked at the Dana-Farber Cancer Institute. For longitudinal analysis, samples were thawed, washed twice in PBS, and stained with anti-human CD45 (PE; BioLegend, 304007) and anti-human CD117 (APC; BioLegend, 313205). CD45lo/CD117+ leukemia cells were FACS-sorted (Sony MA900 sorter) and subsequently processed for RNA-seq (see Methods section on RNA-seq).
Soto-Feliciano Y.M., Sánchez-Rivera F.J., Perner F., Barrows D.W., Kastenhuber E.R., Ho Y.J., Carroll T., Xiong Y., Anand D., Soshnev A.A., Gates L., Beytagh M.C., Cheon D., Gu S., Liu X.S., Krivtsov A.V., Meneses M., de Stanchina E., Stone R.M., Armstrong S.A., Lowe S.W, & Allis C.D. (2022). A Molecular Switch between Mammalian MLL Complexes Dictates Response to Menin–MLL Inhibition. Cancer Discovery, 13(1), 146-169.
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5

Multicolor Flow Cytometry of Immune Cells

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Murine monoclonal antibodies (all IgG1): anti-human CD42b-APC (Biolegend), anti-human CD45-PE (Biolegend), anti-human/mouse C3/C3b/iC3b-FITC (Cedarlane), and anti-human CD11b-PerCP/Cy5.5 (Biolegend). IgG1 isotype controls: APC- and PerCP/Cy5.5-labeled (Biolegend), PE-labeled (Chemicon), FITC-labeled (Cedarlane).
Blatt A.Z., Saggu G., Kulkarni K.V., Cortes C., Thurman J.M., Ricklin D., Lambris J.D., Valenzuela J.G, & Ferreira V.P. (2016). Properdin-mediated C5a production enhances stable binding of platelets to granulocytes in human whole blood. Journal of immunology (Baltimore, Md. : 1950), 196(11), 4671-4680.
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6

Immunophenotyping of GMSCs and Macrophages

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The expression of CD73, CD90, CD105, CD34, CD45, and CD11b in GMSCs and CD11b, CD206, and CD86 in PBMC-derived macrophages were analyzed using the FACS Calibur (Becton Dickinson, USA) and the CellQuest software (Becton Dickinson). The adherent cells were washed with PBS and collected using Accutase (Nacalai Tesque) and resuspended in 50 μL staining buffer (BD Pharmingen, country). The harvested cells were blocked with 2 μL Human Trustain FcX (Fc receptor Blocking Solution; BioLegend) for 10 min at room temperature, and stained with 2 μL antibodies namely, FITC anti-human CD73, FITC anti-human CD90, Alexa Fluor® 488 anti-human CD105, FITC anti-human CD34, PE anti-human CD45, FITC anti-human CD11b, Alexa Fluor® 488 anti-human CD11b, PE anti-human CD206, APC anti-human CD206, PE anti-human CD86 and APC anti-human CD86 (BioLegend) in the dark for 30 min at 4 °C.
Nakao Y., Fukuda T., Zhang Q., Sanui T., Shinjo T., Kou X., Chen C., Liu D., Watanabe Y., Hayashi C., Yamato H., Yotsumoto K., Tanaka U., Taketomi T., Uchiumi T., Le A.D., Shi S, & Nishimura F. (2020). Exosomes from TNF-α-treated human gingiva-derived MSCs enhance M2 macrophage polarization and inhibit periodontal bone loss. Acta biomaterialia, 122, 306-324.
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7

Multiparametric Flow Cytometry Analysis

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Cells were suspended in flow cytometry staining buffer (Invitrogen, San Diego, CA, USA) and filtered into a single-cell suspension with a 40 μm/100 μm mesh. Then, 5 × 105 cells were incubated with FITC-anti-mouse CD86 (1:50), PE-anti-mouse CD206 (1:40), PE-anti-human CD19 (1:40), PE-anti-human CD34 (1:40), PE-anti-human CD11b (1:40), PE-anti-human CD45 (1:40), FITC-anti-human HLA-DR (1:50), PE-anti-human CD73 (1:40), FITC-anti-human CD90 (1:40), and PE-anti-human CD105 (1:40) antibodies (all from BioLegend, San Diego, CA, USA) at 4 °C for 30 min. After washing twice with staining buffer, cells were suspended in 100 μL staining buffer and analyzed via flow cytometry (BD FACS Calibur, Beckman Coulter).
Xu P., Xin Y., Zhang Z., Zou X., Xue K., Zhang H., Zhang W, & Liu K. (2020). Extracellular vesicles from adipose-derived stem cells ameliorate ultraviolet B-induced skin photoaging by attenuating reactive oxygen species production and inflammation. Stem Cell Research & Therapy, 11, 264.
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8

Multicolor Flow Cytometry Analysis

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Pembrolizumab and nivolumab (anti-PD-1) were obtained from Merck and Bristol-Myers Squibb. The following antibodies were used for flow cytometry analyses: FITC anti-human CD4 (BioLegend Cat#391503), PE anti-human CD45 (Biolegend, clone HI30), APC anti-human CD56 (Biolegend, clone 5.1H11), APC/Cyanine7 anti-human CD8a (Biolegend, San Diego, CA, USA, clone RPA-T8), PE/Cyanine7 anti-human CD3 (Biolegend, clone HIT3a), PerCP/Cyanine5.5 anti-human CD314 (NKG2D) (Biolegend, clone 1D11), FITC anti-human CD6 (Biolegend, clone BL-CD6), APC anti-human CD16 (Biolegend, clone 3G8), Pacific Blue anti-human/mouse Ki67 (Biolegend, clone 16A8), FITC anti-human/mouse Granzyme B (Biolegend, clone GB11) and APC anti-human Perforin (Biolegend, clone B-D48).
Gurrea-Rubio M., Wu Q., Amin M.A., Tsou P.S., Campbell P.L., Amarista C.E., Ikari Y., Brodie W.D., Mattichak M.N., Muraoka S., Randon P.M., Lind M.E., Ruth J.H., Mao-Draayer Y., Ding S., Shen X., Cooney L.A., Lin F, & Fox D.A. (2023). Activation of Cytotoxic Lymphocytes Through CD6 Enhances Killing of Cancer Cells. Research Square.
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9

Characterization of MSCs and Macrophages

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The expression levels of cell surface antigens (CD90, CD73, CD45, and CD34) on MSCs and HIF-MSCs and cell surface antigens (CD80, CD86, CD163, and CD206) on macrophages were detected using flow cytometry. Harvested cells were digested with trypsin-EDTA and stained with FITC anti-human CD34, APC anti-human CD73, PE anti-human CD90, PE anti-human CD45, PE anti-mouse CD86, PE anti-human CD163, FITC anti-human CD80 and FITC anti-human CD206 (Biolegend, San Diego, CA, USA), all human-specific antibodies. Immunofluorescence was performed to detect the expression of macrophage surface molecules F4/80, CD86, and CD163 in tissue sections. The expression levels were then analyzed using a BD FACS Calibur cytometer (Beckman, Pasadena, CA, USA). The viability of MSCs was evaluated using trypan blue exclusion and cell counting kit-8 (CCK-8) (Beyotime, Shanghai, China).
Zhu W., Chen Q., Li Y., Wan J., Li J, & Tang S. (2023). HIF-1α-Overexpressing Mesenchymal Stem Cells Attenuate Colitis by Regulating M1-like Macrophages Polarization toward M2-like Macrophages. Biomedicines, 11(3), 825.
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10

Characterization of Immune Cell Phenotypes

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FITC mouse IgG1 κ isotype control (400110), PerCP-Cy5.5 mouse IgG1 κ isotype control (400149), PE mouse IgG1 κ isotype control (400114), PerCP-Cy5.5 anti-human CD3 (300430), FITC anti-human CD8a (300906), PE anti-human CD45 (368510), PerCP-Cy5.5 anti-human IFN-γ (506528), FITC Rat IgG2a κ isotype control (400506), PerCP-Cy5.5 Rat IgG1 κ isotype control (400426), FITC anti-mouse CD3 (100204), FITC anti-mouse CD8a (100705), PE anti-mouse CD45 (103106), and PerCP-Cy5.5 anti-mouse IFN-γ (505822) were purchased from Biolegend. PE anti-human PD-L1 (557924), FITC anti-human PD-L1 (558065), FITC anti-human HLA-A2 (343303), FITC Rat IgG2b κ isotype control (400605), PE Rat IgG2a λ isotype control (400635), and PE anti-mouse PD-L1 (558091) were obtained from BD Biosciences.
Ding L., Chen X., Zhang W., Dai X., Guo H., Pan X., Xu Y., Feng J., Yuan M., Gao X., Wang J., Xu X., Li S., Wu H., Cao J., He Q, & Yang B. (2023). Canagliflozin primes antitumor immunity by triggering PD-L1 degradation in endocytic recycling. The Journal of Clinical Investigation, 133(1), e154754.
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