mouse monoclonal anti osteocalcin antibodies  (R&D Systems)

Journal: iScience

Article Title: cGAS/STING sensing in dendritic cells discriminates between daptomycin sensitive and resistant Staphylococcus aureus clinical isolates

doi: 10.1016/j.isci.2026.115854

Figure Lengend Snippet: TLR2 is required for MutuDC sensing of, but not internalization of MRSA (A) Relative pHrodo labeled MRSA internalization by MutuDC over 4 h following stimulation with DapS A8819 (light blue symbols) or DapR A8817 (dark blue symbols), or media alone (white squares). Prior to stimulation, MutuDC were pre-treated for 1 h with TLR2 blocking antibody (clone T2.5; triangles with dashed lines) or media alone (circles with filled lines). Relative MRSA internalization by each DC subset is expressed as the gMFI of pHrodo. Results show the mean and (SD) of duplicates from one experiment, representative of two independent experiments. (B) Cytokine secretion (pg/mL) by MutuDC stimulated with TLR2 ligand peptidoglycan of S. aureus (PGN-SA) (10 μg/mL) or (C) DapS (A8819; light blue) or DapR (A8817; dark blue) MRSA (MOI of 10) for 18 h. MutuDC were first pre-treated with either TLR2 blocking antibody (dot-filled bars) or media alone (filled bars) as in A, or an isotype control (clone 163D3, empty bars) at 1 μg/mL. Results pooled from four (B) or three (C) independent experiments and expressed as the mean ± SEM, with each symbol (circle, square, and directional triangles) representing paired experimental replicates ( n = 3). Statistical significance determined using paired t test and reported as indicated by an ∗ when p ≤ 0.05. (D) Expression of surface activation markers by MutuDC stimulated with DapS A8819 MRSA. DC were pre-treated with TLR2 blocking antibody (black trace), isotype control (dashed red trace), and media alone (light blue shaded). Unstained control sample is shown for each marker (black dashed trace). Data shown from one experiment, representative of three independent experiments.

Article Snippet: mAB mTLR2- anti-mouse/human TLR2 , InvivoGen , Cat# mab-mtlr2; RRID: AB_763722.

Techniques: Labeling, Blocking Assay, Control, Expressing, Activation Assay, Marker

Journal: Cancers

Article Title: Optimizing Sequential Targeted Therapies in Advanced Renal Cell Carcinoma Using Patient-Derived Orthotopic Xenograft Mouse Avatars

doi: 10.3390/cancers18101615

Figure Lengend Snippet: Histopathological comparison of tumor architecture and proliferation between patient renal cell carcinoma (RCC) specimens and corresponding patient-derived xenograft models. Formalin-fixed, paraffin-embedded sections from primary patient tumors (KiCa-Pt58 and KiCa-Pt118), matched subcutaneous xenografts, and intra-renal patient-derived orthotopic xenograft (PDOX) tumors were analyzed to assess preservation of tumor histology and proliferative characteristics. ( A , C ) Representative images of KiCa-Pt58 ( A ) and KiCa-Pt118 ( C ) tissues stained with hematoxylin and eosin (H&E) for tissue architecture or were subjected to immunohistochemistry (IHC) for human CD44 (tumor cell marker) and human Ki67 (proliferation marker). Brown staining indicates positive immunoreactivity. Images were captured at 100× original magnification using an Axiovert 200M deconvolution microscope and SlideBook 6.0 software (Intelligent Imaging Innovations, Denver, CO, USA). ( B , D ) Quantitative analysis of Ki67-positive area (%) in KiCa-Pt58 ( B ) and KiCa-Pt118 ( D ) tissues. Positive (brown) staining areas were quantified digitally using Adobe Photoshop 7.0, with the percentage of immunoreactive area calculated per field. Data are presented as mean ± SEM (multiple fields per sample). Comparisons among patient biopsy specimens, subcutaneous xenografts, and orthotopic PDOX tumors were performed using unpaired Student’s t -test. No significant differences were observed (ns, p > 0.05), demonstrating faithful recapitulation of the parental tumor proliferative index across model passages. Abbreviations: RCC, renal cell carcinoma; PDOX, patient-derived orthotopic xenograft; H&E, hematoxylin and eosin; IHC, immunohistochemistry.

Article Snippet: Paraffin-embedded sections (5 μm) were stained with H&E or subjected to immunohistochemistry using primary antibodies against human Ki67 (proliferation marker; Thermo Fisher Scientific, Waltham, MA, USA; 1:200), human CD44 (tumor cell marker; Acris Antibodies, Rockville, MD, USA; 1:75), mouse CD31 (angiogenesis marker; Abcam, Cambridge, MA, USA; 1:200), and human PD-L1 (programmed death-ligand 1, immune checkpoint ligand; BioLegend, San Diego, CA, USA; 1:200) [ ].

Techniques: Comparison, Derivative Assay, Formalin-fixed Paraffin-Embedded, Preserving, Staining, Immunohistochemistry, Marker, Microscopy, Software, Imaging

Journal: Cancers

Article Title: Optimizing Sequential Targeted Therapies in Advanced Renal Cell Carcinoma Using Patient-Derived Orthotopic Xenograft Mouse Avatars

doi: 10.3390/cancers18101615

Figure Lengend Snippet: Histopathological and immunohistochemical evaluation of targeted therapy responses in the KiCa-Pt58 patient-derived orthotopic xenograft (PDOX) model. ( A ) Representative images of left kidney tumors from KiCa-Pt58 PDOX mice after vehicle control or sequential targeted therapy (Everolimus→Sunitinib [E→S], Pazopanib→Sunitinib [P→S], Sunitinib→Everolimus [S→E], Pazopanib→Everolimus [P→E]). Formalin-fixed, paraffin-embedded sections were stained with hematoxylin and eosin (H&E) for tumor architecture or subjected to immunohistochemistry (IHC) for human CD44 (tumor cell marker), human Ki67 (proliferation marker), mouse CD31 (angiogenesis/endothelial marker), and human PD-L1 (immune checkpoint ligand). Brown staining indicates positive immunoreactivity. Images were acquired at 100× original magnification using an Axiovert 200M deconvolution microscope and SlideBook 6.0 software (Intelligent Imaging Innovations, Denver, CO, USA). ( B – D ) Quantitative analysis of positive staining area (%) for Ki67 ( B ), CD31 ( C ), and PD-L1 ( D ) across treatment groups. Positive (brown) areas were quantified digitally using Adobe Photoshop 7.0 (percentage immunoreactive area per field). Data are presented as mean ± SEM (multiple fields per sample; n = 7–9 mice per group). Statistical comparisons vs. control were performed using one-way ANOVA followed by Dunnett’s or Tukey’s post hoc tests (GraphPad Prism v7). Asterisks indicate significance: * p < 0.05; ** p < 0.01; *** p < 0.001. Effective regimens (particularly P→E and S→E) significantly reduced Ki67+ proliferation, CD31+ vascularity, and PD-L1 expression compared to control, consistent with antitumor and potential immunomodulatory effects. Abbreviations: PDOX, patient-derived orthotopic xenograft; IHC, immunohistochemistry; E, everolimus; S, sunitinib; P, pazopanib.

Article Snippet: Paraffin-embedded sections (5 μm) were stained with H&E or subjected to immunohistochemistry using primary antibodies against human Ki67 (proliferation marker; Thermo Fisher Scientific, Waltham, MA, USA; 1:200), human CD44 (tumor cell marker; Acris Antibodies, Rockville, MD, USA; 1:75), mouse CD31 (angiogenesis marker; Abcam, Cambridge, MA, USA; 1:200), and human PD-L1 (programmed death-ligand 1, immune checkpoint ligand; BioLegend, San Diego, CA, USA; 1:200) [ ].

Techniques: Immunohistochemical staining, Derivative Assay, Control, Formalin-fixed Paraffin-Embedded, Staining, Immunohistochemistry, Marker, Microscopy, Software, Imaging, Expressing

Journal: Cancers

Article Title: Optimizing Sequential Targeted Therapies in Advanced Renal Cell Carcinoma Using Patient-Derived Orthotopic Xenograft Mouse Avatars

doi: 10.3390/cancers18101615

Figure Lengend Snippet: Histopathological and immunohistochemical assessment of targeted therapy responses in the KiCa-Pt118 patient-derived orthotopic xenograft (PDOX) model. ( A ) Representative images of left kidney tumors from KiCa-Pt118 PDOX mice after vehicle control or sequential targeted therapy (Everolimus→Sunitinib [E→S], Pazopanib→Sunitinib [P→S], Sunitinib→Everolimus [S→E], Pazopanib→Everolimus [P→E]). Formalin-fixed, paraffin-embedded sections were stained with hematoxylin and eosin (H&E) for tumor architecture or subjected to immunohistochemistry (IHC) for human CD44 (tumor cell marker), human Ki67 (proliferation marker), mouse CD31 (angiogenesis/endothelial marker), and human PD-L1 (immune checkpoint ligand). Brown staining indicates positive immunoreactivity. Images were acquired at 100× original magnification using an Axiovert 200M deconvolution microscope and SlideBook 6.0 software (Intelligent Imaging Innovations, Denver, CO, USA). ( B – D ) Quantitative analysis of positive staining area (%) for Ki67 ( B ), CD31 ( C ), and PD-L1 ( D ) across treatment groups. Positive (brown) areas were quantified digitally using Adobe Photoshop 7.0 (percentage immunoreactive area per field), as described in . Data are presented as mean ± SEM (multiple fields per sample; n = 7–9 mice per group). Statistical comparisons vs. control were performed using one-way ANOVA followed by Dunnett’s or Tukey’s post hoc tests (GraphPad Prism v7). Asterisks indicate significance: * p < 0.05; ** p < 0.01; *** p < 0.001. Effective regimens (particularly S→E) significantly reduced Ki67+ proliferation, CD31+ vascularity, and PD-L1 expression compared to control, consistent with antitumor activity in this indolent, non-metastatic model. Abbreviations: PDOX, patient-derived orthotopic xenograft; IHC, immunohistochemistry; E, everolimus; S, sunitinib; P, pazopanib.

Article Snippet: Paraffin-embedded sections (5 μm) were stained with H&E or subjected to immunohistochemistry using primary antibodies against human Ki67 (proliferation marker; Thermo Fisher Scientific, Waltham, MA, USA; 1:200), human CD44 (tumor cell marker; Acris Antibodies, Rockville, MD, USA; 1:75), mouse CD31 (angiogenesis marker; Abcam, Cambridge, MA, USA; 1:200), and human PD-L1 (programmed death-ligand 1, immune checkpoint ligand; BioLegend, San Diego, CA, USA; 1:200) [ ].

Techniques: Immunohistochemical staining, Derivative Assay, Control, Formalin-fixed Paraffin-Embedded, Staining, Immunohistochemistry, Marker, Microscopy, Software, Imaging, Expressing, Activity Assay

Journal: bioRxiv

Article Title: Ablation of glypican-3 enhances radiosensitivity in liver cancer by prolonging G2/M arrest and activating the ATM/CHK2 pathway

doi: 10.64898/2026.05.11.724294

Figure Lengend Snippet: Ectopic GPC3 expression was successfully established and validated in isogenic SNU449 cells. ( A ) SNU449 cells were transduced with hGPC3-mGFP and clonally isolated, and GPC3 mRNA expression was quantified by qPCR. A431/GPC3 and HepG2 cells served as positive controls; A431 and GPC3⁻ HepG2 cells served as negative controls. ( B ) GPC3 protein expression was assessed by Western blotting (GAPDH loading control). ( C ) Cell-surface GPC3 was detected in A431/GPC3, SNU449/GPC3, and HepG2 cells, but not in GPC3⁻ SNU449, SNU449/vector, A431, or HepG2 cells. ( D ) Immunofluorescence staining confirmed GPC3 expression in SNU449/GPC3 and A431/GPC3 cells, but not in parental controls. Scale bars: 50 μ m.

Article Snippet: Human tissue specimens were incubated with a mouse monoclonal anti-human GPC3 antibody (clone 1G12; Cell Marque) under the same conditions.

Techniques: Expressing, Transduction, Isolation, Western Blot, Control, Plasmid Preparation, Immunofluorescence, Staining

Journal: bioRxiv

Article Title: Ablation of glypican-3 enhances radiosensitivity in liver cancer by prolonging G2/M arrest and activating the ATM/CHK2 pathway

doi: 10.64898/2026.05.11.724294

Figure Lengend Snippet: GPC3 expression regulates tumor cell proliferation and radiosensitivity. Proliferation of paired GPC3-positive (GPC3⁺) and GPC3-deficient (GPC3⁻) HepG2 ( A ), Hep3B ( B ), SNU449 and SNU449/V (empty vector control) ( C ), and A431 ( D ) cells was monitored using the IncuCyte® live-cell analysis system under non-irradiated (Non-IR) conditions and following 6 Gy irradiation (IR). Phase-area confluence was normalized to 0 h, and differences in proliferation kinetics were analyzed by two-way ANOVA, with significance assessed at the final time point (****, P <0.0001). Radiosensitivity was evaluated by clonogenic survival assays in the corresponding GPC3⁺ and GPC3⁻ HepG2 ( E ), Hep3B ( F ), SNU449 ( G ), and A431 ( H ) cells exposed to graded doses of γ-irradiation. Colonies were quantified 10–14 days later to generate survival curves. GPC3 loss increased radiosensitivity, yielding dose-modifying factors (DMFs) of 1.25 (HepG2), 1.36 (Hep3B), 1.20 (SNU449), and 1.75 (A431). Data represent mean surviving fraction ± SD from more than three independent experiments.

Article Snippet: Human tissue specimens were incubated with a mouse monoclonal anti-human GPC3 antibody (clone 1G12; Cell Marque) under the same conditions.

Techniques: Expressing, Plasmid Preparation, Control, Cell Analysis, Irradiation

Journal: bioRxiv

Article Title: Ablation of glypican-3 enhances radiosensitivity in liver cancer by prolonging G2/M arrest and activating the ATM/CHK2 pathway

doi: 10.64898/2026.05.11.724294

Figure Lengend Snippet: GPC3 deficiency increases IR-induced DNA double-strand breaks in tumor cells. DNA damage responses were assessed in paired GPC3-positive (GPC3⁺) and GPC3-deficient (GPC3⁻) HepG2 ( A , E ), Hep3B ( B , F ), SNU449 ( C , G ), and A431 ( D , I ) cells following 6 Gy irradiation (IR). γ-H2AX immunocytochemistry ( A – D ) was performed at the indicated time points to quantify DNA double-strand break formation and resolution. Representative images at control, 1 h, and 48 h post-IR are shown. γ-H2AX foci (magenta) were quantified from > 50 cells per condition; nuclei were counterstained with DAPI (blue). Scale bars: 25 μ m. Statistical comparisons of foci numbers were performed using a t -test (* p <0.05; *** p <0.001). Comet assays ( E – I ) were performed 24 h after IR. Representative comet images (left) and quantified tail moments (right) are shown. Data are presented as mean ± SD, and significance was determined using a t -test (** p <0.01; *** p <0.001; **** p <0.0001). Scale bars: 50 μ m.

Article Snippet: Human tissue specimens were incubated with a mouse monoclonal anti-human GPC3 antibody (clone 1G12; Cell Marque) under the same conditions.

Techniques: Irradiation, Immunocytochemistry, Control

Journal: bioRxiv

Article Title: Ablation of glypican-3 enhances radiosensitivity in liver cancer by prolonging G2/M arrest and activating the ATM/CHK2 pathway

doi: 10.64898/2026.05.11.724294

Figure Lengend Snippet: GPC3 deficiency enhances radiation response in xenograft models and correlates with clinical outcomes. ( A – D ) HepG2/Luc xenografts derived from GPC3⁺ or GPC3⁻ cells with or without 10 Gy irradiation were monitored by bioluminescence imaging (BLI) and tumor volume measurements. GPC3⁻ tumors showed enhanced growth delay following irradiation. Representative GPC3 and Ki-67 staining is shown (scale bar: 100 μ m). ( E – H ) Similar analyses in A431/Luc xenografts demonstrated greater radiation sensitivity in GPC3⁻ tumors. ( I ) In a clinical cohort, high tumor GPC3 expression was associated with poorer overall survival ( p = 0.008). Statistical comparisons were performed by two-way ANOVA; significance was assessed at the final time point (* p <0.05; *** p <0.001; **** p <0.0001).

Article Snippet: Human tissue specimens were incubated with a mouse monoclonal anti-human GPC3 antibody (clone 1G12; Cell Marque) under the same conditions.

Techniques: Derivative Assay, Irradiation, Imaging, Staining, Expressing