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Anti human cd45

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Anti-human-CD45 is a monoclonal antibody used for the identification and enumeration of human leukocytes. It binds to the CD45 antigen, which is expressed on the surface of all human leukocytes, including T cells, B cells, monocytes, granulocytes, and natural killer cells.

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Lab products found in correlation

Apc annexinv apoptosis kit 1, by BD (2 mentions) Foxp3 transcription factor staining buffer set, by Thermo Fisher Scientific (2 mentions) Propidium iodide, by Merck Group (2 mentions) Cell proliferation kit 1, by Roche (2 mentions) Cell staining buffer, by BioLegend (1 mentions) Gentle cell dissociation reagent (gcdr), by STEMCELL (1 mentions) Facscanto 2, by BD (1 mentions) Oxidized ldl, by Thermo Fisher Scientific (1 mentions) C57bl 6, by Jackson ImmunoResearch (1 mentions) Isotype control, by Thermo Fisher Scientific (1 mentions) Anti human cd29, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Anti-human CD45 (clone 2D1, Anti-human CD45 APC Anti-human CD45 (clone HI30; Anti-human CD45-FitC PerCP/Cy5.5 anti-human CD45 Anti-Human CD45 PE Mouse anti-human CD45-FITC FITC-conjugated mouse anti-human CD45 Anti-CD45

5 protocols using anti human cd45

1

Combinatorial Cytotoxicity and Cell Cycle Regulation

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We analyzed cell line viability upon treatment with BPTES (10 μM) and DBZ (250 nM) alone and in combination using the Cell Proliferation Kit I (Roche). We analyzed apoptosis by flow cytometry with APC AnnexinV Apoptosis Kit I (BD Biosciences). We used Propidium Iodide (Sigma) DNA staining to analyze cell cycle distribution.
For the metabolic rescue experiment, cells were maintained on RPMI buffered with 40 mM HEPES, and treated with methyl pyruvate (10 mM) or dimethyl ketoglutarate (8 mM) and/or DBZ (250 nM). We added new media every two days.
For the measurement of PTEN intracellular levels in PDTALL 19 human primary xenografted samples, we removed red cells from peripheral blood samples by incubation with red blood cell lysis buffer (155 mM NH4Cl, 12 mM KHCO3 and 0.1 mM EDTA) for 5 min at room temperature. We then stained the samples with anti-human-CD45 (eBioscience, Cat# 17-0459-42, clone HI30, dilution 1:200). Next, we performed fixation/permeabilization using Foxp3/Transcription Factor Staining Buffer Set (eBioscience Cat# 00-5523), following manufacturer’s instructions. Finally, we performed intracellular staining with anti-PTEN-PE (BD Phosflow, Cat# 560002, clone A2B1, dilution 1:5) and we used PE-labeled mouse IgG1 isotype as a control (BD Pharmingen, Cat# 551436, dilution 1:5).
Herranz D., Ambesi-Impiombato A., Sudderth J., Sánchez-Martín M., Belver L., Tosello V., Xu L., Wendorff A.A., Castillo M., Haydu J.E., Márquez J., Matés J.M., Kung A.L., Rayport S., Cordon-Cardo C., DeBerardinis R.J, & Ferrando A.A. (2015). Metabolic reprogramming induces resistance to anti-NOTCH1 therapies in acute lymphoblastic leukemia. Nature medicine, 21(10), 1182-1189.
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2

Combinatorial Cytotoxicity and Cell Cycle Regulation

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We analyzed cell line viability upon treatment with BPTES (10 μM) and DBZ (250 nM) alone and in combination using the Cell Proliferation Kit I (Roche). We analyzed apoptosis by flow cytometry with APC AnnexinV Apoptosis Kit I (BD Biosciences). We used Propidium Iodide (Sigma) DNA staining to analyze cell cycle distribution.
For the metabolic rescue experiment, cells were maintained on RPMI buffered with 40 mM HEPES, and treated with methyl pyruvate (10 mM) or dimethyl ketoglutarate (8 mM) and/or DBZ (250 nM). We added new media every two days.
For the measurement of PTEN intracellular levels in PDTALL 19 human primary xenografted samples, we removed red cells from peripheral blood samples by incubation with red blood cell lysis buffer (155 mM NH4Cl, 12 mM KHCO3 and 0.1 mM EDTA) for 5 min at room temperature. We then stained the samples with anti-human-CD45 (eBioscience, Cat# 17-0459-42, clone HI30, dilution 1:200). Next, we performed fixation/permeabilization using Foxp3/Transcription Factor Staining Buffer Set (eBioscience Cat# 00-5523), following manufacturer’s instructions. Finally, we performed intracellular staining with anti-PTEN-PE (BD Phosflow, Cat# 560002, clone A2B1, dilution 1:5) and we used PE-labeled mouse IgG1 isotype as a control (BD Pharmingen, Cat# 551436, dilution 1:5).
Herranz D., Ambesi-Impiombato A., Sudderth J., Sánchez-Martín M., Belver L., Tosello V., Xu L., Wendorff A.A., Castillo M., Haydu J.E., Márquez J., Matés J.M., Kung A.L., Rayport S., Cordon-Cardo C., DeBerardinis R.J, & Ferrando A.A. (2015). Metabolic reprogramming induces resistance to anti-NOTCH1 therapies in acute lymphoblastic leukemia. Nature medicine, 21(10), 1182-1189.
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3

Characterization of Immune Infiltrates in Organoids

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A digestion step was performed for the harvest and characterization of lymphocytes infiltrated into the organoids. Briefly, organoids were removed from culture insert after immune infiltration experiments, washed with PBS twice to eliminate the PBMCs, which are not inside the organoids, and incubated with Gentle Cell Dissociation reagent (Stem Cell Technologies; catalog no. 07174) for 2 min at RT. Next, the organoids were mechanically disrupted by pipetting. After complete disaggregation, single cell suspensions were dissolved in Cell Staining buffer (Biolegend, catalog no. 420201) and incubated at 4°C for 10 min with antibody staining.
Cells were stained with anti-human CD45 (eBioscience; catalog no. 17-0459-42), CD3 (Biolegend; catalog no. 344840), CD8 (Biolegend; catalog no. 344709), CD4 (Biolegend; catalog no. 300505), CD56 (BD Pharmingen; catalog no. 556647) and CD19 (Biolegend; catalog no. 363005) antibodies. Cell viability was assessed by negative live/dead antibody staining (Miltenyi Biotech; catalog no. 130-109-816). Analysis was performed on a BD FACS Canto II (BD Biosciences). BV421+ and single cells were gated out, followed by CD45+ cell selection. Analysis of CD3, CD8, CD4, CD56, and CD19 lymphocyte populations was performed with FlowJo software (Tree Star Inc.)
Soldi R., Ghosh Halder T., Weston A., Thode T., Drenner K., Lewis R., Kaadige M.R., Srivastava S., Daniel Ampanattu S., Rodriguez del Villar R., Lang J., Vankayalapati H., Weissman B., Trent J.M., Hendricks W.P, & Sharma S. (2020). The novel reversible LSD1 inhibitor SP-2577 promotes anti-tumor immunity in SWItch/Sucrose-NonFermentable (SWI/SNF) complex mutated ovarian cancer. PLoS ONE, 15(7), e0235705.
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4

Characterization of DPP4 and CD36 Expression in Murine Immune Cells

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C57BL/6, MyD88−/−, TRIF−/−, and CD36−/− mice were purchased from Jackson Laboratory. All procedures were approved by the IACUC committee at the Case Western Reserve University.
The antibodies used for flow cytometry were purchased from the following companies: anti-human DPP4 (clone # 2A6 [PE-labeled], purchased from eBioscience, San Diego, CA; Clone # BA5b [APC-labeled], purchased from Biolegend, San Diego, CA), PE-labeled anti-mouse DPP4 (Clone # 155202, R&D system, Minneapolis, MN), anti-human CD36 (clone # 5–271 [PE- or APC-labeled], Biolegend, San Diego, CA), APC-labeled anti-mouse CD36 (clone # 72–1, eBioscience, San Diego, CA), PE/Cy5-labeled anti-human/mouse CD11b (Clone # M1/70, Biolegend, San Diego, CA), FITC-labeled anti-human CD3 (Clone # OKT3, eBioscience, San Diego, CA), anti-human CD45 (Clone # Hi30, eBioscience, San Diego, CA), and anti-human CD4 (Clone # OKT4, eBioscience, San Diego, CA). Oxidized LDL was purchased from Thermo Fisher Scientific (Cat. # AAJ652618PL, Thermo Fisher Scientific, Waltham, MA). DPP4 enzymatic inhibitor (DPP4i) Linagliptin was a kind gift from Boehringer Ingelheim (Ingelheim am Rhein, Germany).
Rao X., Zhao S., Braunstein Z., Mao H., Razavi M., Duan L., Wei Y., Toomey A.C., Rajagopalan S, & Zhong J. (2019). Oxidized LDL upregulates macrophage DPP4 expression via TLR4/TRIF/CD36 pathways. EBioMedicine, 41, 50-61.
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5

Phenotyping of hUC-MSCs by Flow Cytometry

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In order to perform flow cytometry analysis, the 3rd to 5th passage of 3 lines of hUC-MSCs were dissociated into single cells by TrypLE Express and followed by resuspended 1 × 105 hUC-MSCs in 100 μl of 1 mg/ml bovine serum albumin (BSA)/DPBS (Thermo Fisher Scientific). Cells were incubated with the following allophycocyanin (APC)-conjugated antibodies in dark for one hour, which included antihuman CD34, antihuman CD45, antihuman CD29, antihuman CD73, antihuman CD90, antihuman CD105, antihuman CD44, antihuman HLA-DR, and isotype controls (eBioscience, San Diego, California, USA). Cells were washed twice with ice cold 1 mg/ml BSA/DPBS and resuspended in 400 μl of 1 mg/ml BSA/DPBS. Subsequently, these surface markers were evaluated by flow cytometer (BD Biosciences, San Jose, California, USA). Data was analyzed using FlowJo software (BD Biosciences).
Zhou S., Lei Y., Wang P., Chen J., Zeng L., Qu T., Maldonado M., Huang J., Han T., Wen Z., Tian E., Meng X., Zhong Y, & Gu J. (2022). Human Umbilical Cord Mesenchymal Stem Cells Encapsulated with Pluronic F-127 Enhance the Regeneration and Angiogenesis of Thin Endometrium in Rat via Local IL-1β Stimulation. Stem Cells International, 2022, 7819234.
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