anti hcd45 fitc  (Thermo Fisher)

Journal: bioRxiv

Article Title: Profiling histone post-translational modifications to identify signatures of epigenetic drug response in T-cell acute lymphoblastic leukemia

doi: 10.1101/2025.09.01.673463

Figure Lengend Snippet: (A) NSG mice were engrafted with malignant primary cells derived from the bone marrow of 10 T-ALL patients for tumor expansion. Upon disease establishment, mice were sacrificed, and leukemic cells were harvested from the spleen. To mimic biological replicates, six mice were injected per patient. The percentage of human CD45+ cells (hCD45%) was measured by flow cytometry, with only samples exceeding 85% used for downstream analysis. Global hPTM profiling and DNA methylation sequencing were performed for all 10 PDX models. In parallel, PDX cells were treated ex vivo with a panel of eight epidrugs to assess drug sensitivity. Spearman correlation analysis was then conducted to explore the relationship between hPTM levels and drug response. Figure created with Biorender.com . (B) Clustered and annotated heatmap displaying the normalized and summarized single hPTM levels across the 10 PDX models. (C) Volcano plot illustrating the differential analysis of hPTMs between CIMPlow and CIMPhigh PDX models.

Article Snippet: Lymphoblast engraftment and leukemic burden was monitored by flow cytometric analysis on peripheral blood, using human CD45+ (hCD45) staining (CD45-FITC antibody; Miltenyi Biotec, Bergish Gladbag, Germany).

Techniques: Derivative Assay, Injection, Flow Cytometry, DNA Methylation Assay, Sequencing, Ex Vivo

Journal: Haematologica

Article Title: Targeting hyperactive platelet-derived growth factor receptor-β signaling in T-cell acute lymphoblastic leukemia and lymphoma

doi: 10.3324/haematol.2023.283981

Figure Lengend Snippet: Selective inhibition of PDGFRB in vivo results in significant leukemia growth delay. (A) % hCD45 + cells in peripheral blood at start of treatment (week 0), 1 and 2 weeks after start of treatment analyzed using flow cytometry. (B) % hCD45 + cells in bone marrow at end of treatment (2 weeks) analyzed using flow cytometry. (C) Spleen weight at end of treatment (2 weeks). ** P <0.01, **** P <0.0001.

Article Snippet: At regular timepoints, % hCD45 (130-114-569, Miltenyi Biotec) was measured using flow cytometry (BD LSR II Flow Cytometer) in peripheral blood (PB).

Techniques: Inhibition, In Vivo, Flow Cytometry

Journal: PLoS ONE

Article Title: Anti-Proteinase 3 Anti-Neutrophil Cytoplasm Autoantibodies Recapitulate Systemic Vasculitis in Mice with a Humanized Immune System

doi: 10.1371/journal.pone.0028626

Figure Lengend Snippet: Characterization of the chimeric immune system.

Article Snippet: Leukocytes were labelled with the following antibodies all at 1∶200 dilution: anti-hCD45-FITC, or –PE, hCD3-FITC, anti-hCD56-Al488 anti-hCD16-PE, anti-CD11b-APC, -PE or -FITC, anti-mCD45-FITC or -PE anti-mB220-APC (EBioscience), anti-hCD14-Alexa647, anti-hCD19-APC, anti-hCD15-FITC, anti-hCD66b-FITC (Biolegend), anti-Ly6C-FITC (BD Pharmingen).

Techniques:

Journal: PLoS ONE

Article Title: Anti-Proteinase 3 Anti-Neutrophil Cytoplasm Autoantibodies Recapitulate Systemic Vasculitis in Mice with a Humanized Immune System

doi: 10.1371/journal.pone.0028626

Figure Lengend Snippet: (A–D) Flow cytometric analysis of leukocytes from tail bleeds six weeks after administration of HSCs (n = 26 mice). (A) Plots showing mouse leukocytes labelled with anti-mouse CD45 antibodies. Compared with control wild-type mouse blood, chimeras have populations of mCD45 negative leukocytes that show SSC characteristics of granulocytes (High), monocytes (Int) and lymphocytes (low). (B) Chimera blood leukocytes express human CD45 and many of these express CD11b. hCD45+,CD11b+ leukocytes predominantly express hCD15 and hCD66b compared with hCD45+,CD11b− leukocytes shown in histograms. (C) A proportion of hCD45+ leukocytes express CD19. (D) Some hCD45 leukocytes are CD14 high and some are CD16+,CD14 low . (E) In chimera bone marrow there are CD11b+ leukocytes which do not express mCD45 and among hCD45+ leukocytes a proportion express CD14 and a proportion express CD66b. (F) In chimera spleen there are CD11b+ leukocytes which express hCD45 and among hCD45+ leukocytes many express both CD14 and CD16. (G) Bone marrow spreads from wild type or chimera mice, labelled with anti-hMPO or anti-hPR3 IgG antibodies (red) purified from patients with vasculitis. Note that chimera bone marrow demonstrates anti-hMPO or anti-hPR3 antibody positive leukocytes with characteristic human neutrophil nuclear morphology. Wild type mouse bone marrow shows no cells positive for these antigens indicating that the anti-human antibodies do not cross react with mouse neutrophils.

Article Snippet: Leukocytes were labelled with the following antibodies all at 1∶200 dilution: anti-hCD45-FITC, or –PE, hCD3-FITC, anti-hCD56-Al488 anti-hCD16-PE, anti-CD11b-APC, -PE or -FITC, anti-mCD45-FITC or -PE anti-mB220-APC (EBioscience), anti-hCD14-Alexa647, anti-hCD19-APC, anti-hCD15-FITC, anti-hCD66b-FITC (Biolegend), anti-Ly6C-FITC (BD Pharmingen).

Techniques: Purification

Journal: PLoS ONE

Article Title: Anti-Proteinase 3 Anti-Neutrophil Cytoplasm Autoantibodies Recapitulate Systemic Vasculitis in Mice with a Humanized Immune System

doi: 10.1371/journal.pone.0028626

Figure Lengend Snippet: Kidney sections were incubated with anti-mCD45 (red) and anti-hCD45 (green) antibodies and images were captured by fluorescence microscopy (T = tubule). Occasional (<5%) glomeruli of anti-PR3 treated mice displayed intense extracapillary leukocyte infiltration (A) in the shape of crescents (arrows). Most glomeruli in animals treated with anti-PR3 antibodies (n = 18) had evidence of intraglomerular (B,G) and peri-glomerular (C,G) leukocyte infiltration. These were comprised mostly of mCD45+ cells, although some hCD45 leukocytes were also present (arrowheads). In addition, there was a significant increase in peri-vascular leukocyte (mCD45+ and hCD45+) infiltration in anti-PR3 treated mice (D,G [per arteriolar section (art.sec.)]). Sections were also stained for deposition of IgG [red] (E,G) and C3 [green] (F,G). IgG was detectable within periglomerular cells, but there was minimal deposition within the glomeruli. Mouse C3 was weakly deposited in glomeruli but was no different between control group (n = 8) and anti-PR3 group (n = 18). Note mouse C3 can be detected normally binding avidly to tubular basement membranes. (Marker = 10 µm) (* P <0.05, ** P <0.01. median ± IQ ± max/min values). (H) Kidney sections from anti-PR3 and control treated animals were incubated with anti-PR3 positive ANCA IgG. In the peritubular capillaries of chimera mice that received anti-PR3 hIgG occasional leukocytes detected by anti-hPR3 hIgG could be detected. No positively stained human neutrophils were seen in glomeruli.

Article Snippet: Leukocytes were labelled with the following antibodies all at 1∶200 dilution: anti-hCD45-FITC, or –PE, hCD3-FITC, anti-hCD56-Al488 anti-hCD16-PE, anti-CD11b-APC, -PE or -FITC, anti-mCD45-FITC or -PE anti-mB220-APC (EBioscience), anti-hCD14-Alexa647, anti-hCD19-APC, anti-hCD15-FITC, anti-hCD66b-FITC (Biolegend), anti-Ly6C-FITC (BD Pharmingen).

Techniques: Incubation, Fluorescence, Microscopy, Staining, Binding Assay, Marker