human b7-h3 antibody  (Bio-Techne corporation)

Journal: bioRxiv

Article Title: Transcription Factor–Mediated Reprogramming of Cancer-Associated Fibroblasts Reveals Targetable Vulnerabilities in Solid Tumors

doi: 10.64898/2026.04.15.718753

Figure Lengend Snippet: A. Immunocytochemistry (ICC) analysis demonstrates strong B7-H3 (CD276) expression on both PC-3 cancer cells and pCAFs, visualized by red fluorescence. B. In 2D co-culture assays, anti-B7-H3 CAR T cells exhibit antigen-specific cytotoxicity against PC-3 cells, with killing efficiency increasing across graded effector-to-target (E:T) ratios.

Article Snippet: Primary human anti–B7-H3 antibody (AF1027, R&D Systems) was diluted in cold staining buffer (0.5 μg/μL; 2 μL per 100 μL reaction volume) and applied to cells for 20–30 min at 4 °C.

Techniques: Immunocytochemistry, Expressing, Fluorescence, Co-Culture Assay

Journal: bioRxiv

Article Title: Transcription Factor–Mediated Reprogramming of Cancer-Associated Fibroblasts Reveals Targetable Vulnerabilities in Solid Tumors

doi: 10.64898/2026.04.15.718753

Figure Lengend Snippet: Stromal Effects on CAR T-Cell Activity in 3D-Bioprinted PC-3 Spheroids. A. Representative images of stromal populations influencing anti-B7-H3 CAR T-cell killing function . B. PC-3 spheroid with pCAF/rpNFs interaction with CAR-T cells monitored by IncuCyte imaging system. C. The killing effects of B7-H3 CAR T cells on PC-3 cells in day 5 compared to day 1. D . Imaging of live cells at different time.

Article Snippet: Primary human anti–B7-H3 antibody (AF1027, R&D Systems) was diluted in cold staining buffer (0.5 μg/μL; 2 μL per 100 μL reaction volume) and applied to cells for 20–30 min at 4 °C.

Techniques: Activity Assay, Imaging

Journal: Advanced healthcare materials

Article Title: Detection of Extracellular Vesicles with Colocalized Surface Markers via a Capture–Release–Capture Strategy for Treatment Monitoring in Ewing Sarcoma

doi: 10.1002/adhm.202505917

Figure Lengend Snippet: Schematic of the ES EV CaReCa Assay for Quantifying ES EVs Displaying Colocalized CD99 and B7-H3 in Patient Plasma. The two-step sequential enrichment workflow introduces three layers of molecular specificity that suppress background EV signals. Step 1 : DTB-grafted anti-CD99 labels CD99 + ES EVs in plasma, which are Ca ptured onto SA-coated Dynabeads (first layer of specificity). Prior to release, TCO-grafted anti-B7-H3 antibodies are added to label CD99 + /B7-H3 + EVs. Exposure to biotin competitively displaces DTB, Re leasing the dual-labeled EVs into solution (second layer of specificity). Step 2 : The TCO-labeled CD99 + /B7-H3 + EVs undergo a bioorthogonal click reaction with mTz-coated EV Click MagBeads, selectively Ca pturing EVs displaying both CD99 and B7-H3 (third layer of specificity). The enriched CD99 + /B7-H3 + ES EVs are then lysed, and encapsulated ACTB mRNA (a stable housekeeping transcript) is quantified by RT-dPCR to determine ES EV abundance. ES, Ewing Sarcoma; EV, Extracellular Vesicle; CaReCa, Capture–Release–Capture; DTB, desthiobiotin; SA, streptavidin; TCO, trans-cyclooctene; mTz, methyltetrazine; ACTB , β -actin; RT-dPCR, reverse transcription digital droplet PCR.

Article Snippet: Hematoxylin and eosin (H&E) staining and IHC were performed on a Ventana Benchmark ULTRA automated system using validated and optimized protocols for anti-human CD99 and anti-human B7-H3 antibodies (R&D Systems).

Techniques: Clinical Proteomics, Labeling, Reverse Transcription

Journal: Advanced healthcare materials

Article Title: Detection of Extracellular Vesicles with Colocalized Surface Markers via a Capture–Release–Capture Strategy for Treatment Monitoring in Ewing Sarcoma

doi: 10.1002/adhm.202505917

Figure Lengend Snippet: Validation of Paired Surface Markers, CD99 and B7-H3, using ES TMA and A673 ES Cells. (A) Representative H&E staining shows ES tissue morphology, while IHC staining demonstrates strong membrane expression of CD99 and B7-H3 on ES TMA slides. Scale bars = 100 μm. (B) Heatmap and pie charts (CD99, n = 57; B7-H3, n = 47) summarizing IHC staining intensities as strong (3+), moderate (2+), or weak (1+); pie charts exclude detached TMA samples. (C) Representative IF images of A673 ES cells showing colocalized membrane expression of CD99 and B7-H3, with nuclei counterstained by DAPI. Blue: DAPI; green: CD99; red: B7-H3; Schematic illustration depicts antibody binding on the ES cell surface. Scale bar = 10 μm. ES, Ewing Sarcoma; TMA, tissue microarray; H&E, hematoxylin and eosin; IHC, immunohistochemistry; ICC, immunocytochemistry; DAPI, 4′,6-diamidino-2-phenylindole.

Article Snippet: Hematoxylin and eosin (H&E) staining and IHC were performed on a Ventana Benchmark ULTRA automated system using validated and optimized protocols for anti-human CD99 and anti-human B7-H3 antibodies (R&D Systems).

Techniques: Biomarker Discovery, Staining, Immunohistochemistry, Membrane, Expressing, Binding Assay, Microarray, Immunocytochemistry

Journal: Advanced healthcare materials

Article Title: Detection of Extracellular Vesicles with Colocalized Surface Markers via a Capture–Release–Capture Strategy for Treatment Monitoring in Ewing Sarcoma

doi: 10.1002/adhm.202505917

Figure Lengend Snippet: Characterization of A673-Derived ES EVs Immobilized via DTB- and Click Chemistry-Mediated Enrichment. (A) Schematic of independent enrichment of A673 ES EVs using DTB-grafted anti-CD99 with SA–Dynabeads and TCO-grafted anti-B7-H3 with EV Click MagBeads, followed by immunogold staining for CD63 or B7-H3 to confirm EV identity and marker colocalization. (B) SEM images showing immobilization of CD99 + and B7-H3 + ES EVs on SA–Dynabeads and EV Click MagBeads, respectively. Scale bars = 1 μm. (C) TEM validation of EV marker expression and colocalization: CD63 (canonical EV marker) was detected on both CD99 + and B7-H3 + EVs using 12 nm nanogold-conjugated antibodies, while B7-H3 was additionally confirmed on CD99 + EVs, establishing the presence of CD99 + /B7-H3 + ES EVs. Scale bars = 100 nm. ES, Ewing Sarcoma; EV, Extracellular Vesicle; DTB, desthiobiotin; SA, streptavidin; TCO, trans-cyclooctene; SEM, scanning electron microscopy; TEM, transmission electron microscopy.

Article Snippet: Hematoxylin and eosin (H&E) staining and IHC were performed on a Ventana Benchmark ULTRA automated system using validated and optimized protocols for anti-human CD99 and anti-human B7-H3 antibodies (R&D Systems).

Techniques: Derivative Assay, Staining, Marker, Biomarker Discovery, Expressing, Electron Microscopy, Transmission Assay

Journal: Advanced healthcare materials

Article Title: Detection of Extracellular Vesicles with Colocalized Surface Markers via a Capture–Release–Capture Strategy for Treatment Monitoring in Ewing Sarcoma

doi: 10.1002/adhm.202505917

Figure Lengend Snippet: Diagnostic Performance of ES EV CaReCa Assay. (A) Schematic workflow of the two-step CaReCa assay applied to plasma (100 μL) from ES patients ( n = 20) and HD ( n = 20). Enriched CD99 + /B7-H3 + ES EVs were lysed, and ACTB mRNA copy numbers were quantified by RT-dPCR. (B) Heatmap of dPCR readouts showing elevated levels of CD99 + /B7-H3 + ES EVs in ES patients compared with HDs. (C) Box plot showing significantly higher dPCR readouts in ES vs. HDs ( p < 0.0001). (D) ROC curve demonstrating high diagnostic performance (AUROC = 0.98) for distinguishing ES from HDs. An unpaired Student’s t-test was used for statistical analysis between ES and HD groups. ES, Ewing Sarcoma; EV, Extracellular Vesicle; CaReCa, Capture–Release–Capture; ACTB , β -actin; RT-dPCR, reverse transcription digital droplet PCR; HD, Healthy Donor; ROC, Receiver Operating Characteristic; AUROC, Area Under the Receiver Operating Characteristic Curve.

Article Snippet: Hematoxylin and eosin (H&E) staining and IHC were performed on a Ventana Benchmark ULTRA automated system using validated and optimized protocols for anti-human CD99 and anti-human B7-H3 antibodies (R&D Systems).

Techniques: Diagnostic Assay, Clinical Proteomics, Reverse Transcription

Journal: Advanced healthcare materials

Article Title: Detection of Extracellular Vesicles with Colocalized Surface Markers via a Capture–Release–Capture Strategy for Treatment Monitoring in Ewing Sarcoma

doi: 10.1002/adhm.202505917

Figure Lengend Snippet: Longitudinal monitoring of ES patients using the ES EV CaReCa assay. Serial plasma samples were analyzed to quantify ACTB mRNA copy numbers in CaReCa-enriched CD99 + /B7-H3 + ES EVs, and results were compared with PET/CT imaging during treatment and surveillance. (A) In a patient with progressive disease, dPCR readouts fluctuated over the diagnostic cutoff during chemotherapy but rose markedly after treatment completion, aligning with PET/CT evidence of metastatic spread to the brain and spine. (B) In a patient with stable disease, dPCR readouts remained below the cutoff throughout the therapy, consistent with PET/CT findings showing sustained response, successful surgery, and no relapse. ES, Ewing Sarcoma; EV, Extracellular Vesicle; CaReCa, Capture–Release–Capture; ACTB , β -actin; PET/CT, Positron Emission Tomography/Computed Tomography.

Article Snippet: Hematoxylin and eosin (H&E) staining and IHC were performed on a Ventana Benchmark ULTRA automated system using validated and optimized protocols for anti-human CD99 and anti-human B7-H3 antibodies (R&D Systems).

Techniques: Clinical Proteomics, Positron Emission Tomography-Computed Tomography, Imaging, Diagnostic Assay, Positron Emission Tomography, Computed Tomography

Journal: Frontiers in Oncology

Article Title: Association of B7H3 with HIF-1α nuclear expression indicates poor prognosis and therapeutic potential in gastric cancer

doi: 10.3389/fonc.2026.1744341

Figure Lengend Snippet: (A) Analysis of B7H3 and HIF-1α gene expression levels in gastric cancer (GC) and adjacent noncancerous tissues. (B) Analysis of the correlation between B7H3 and HIF-1α in gastric adenocarcinoma using the GEPIA database. (C) Analysis of the correlation between B7H3 and HIF-1α in adjacent noncancerous tissues using the GEPIA database. (D) Survival curve of patients with gastric cancer based on B7H3 expression. (E) Survival curve of patients with gastric cancer based on HIF-1α expression. (F) Survival curve of patients with gastric cancer based on combined B7H3 and HIF-1α expression. The red line indicates the high expression group, the blue line indicates the low expression group, and the yellow line indicates the other groups. **** indicates P < 0.001.

Article Snippet: Subsequently, the sections were incubated overnight at 4°C with mouse anti-human B7H3 antibody (R&D Systems, MN, USA, #AF1027, 1:200) or rabbit anti-human HIF-1α antibody(R&D Systems, MN, USA, #MAB19352,Clone #2443C,1:200).

Techniques: Gene Expression, Expressing

Journal: Frontiers in Oncology

Article Title: Association of B7H3 with HIF-1α nuclear expression indicates poor prognosis and therapeutic potential in gastric cancer

doi: 10.3389/fonc.2026.1744341

Figure Lengend Snippet: (A) Representative immunohistochemical (IHC) images of B7H3 and HIF-1α expression in gastric cancer tissues from 268 clinical patients with gastric cancer. (B) Representative IHC images of B7H3 and HIF-1α nuclear expression in gastric cancer tissues from 268 clinical patients with gastric cancer. Scale bars: 100 μm; 25 μm. (C) resentative multiplex fluorescence immunohistochemistry (mIHC) images of high B7H3 expression and HIF-1α nuclear expression in gastric cancer tissues. Scale bar: 100 μm.

Article Snippet: Subsequently, the sections were incubated overnight at 4°C with mouse anti-human B7H3 antibody (R&D Systems, MN, USA, #AF1027, 1:200) or rabbit anti-human HIF-1α antibody(R&D Systems, MN, USA, #MAB19352,Clone #2443C,1:200).

Techniques: Immunohistochemical staining, Expressing, Multiplex Assay, Fluorescence, Immunohistochemistry

Journal: Frontiers in Oncology

Article Title: Association of B7H3 with HIF-1α nuclear expression indicates poor prognosis and therapeutic potential in gastric cancer

doi: 10.3389/fonc.2026.1744341

Figure Lengend Snippet: (A) Survival curve of gastric cancer patients based on B7H3 expression levels. (B) Survival curve of patients with gastric cancer based on HIF-1α expression levels. (C) Survival curve of patients with gastric cancer based on whether HIF-1α was expressed in the nucleus. (D) Survival curve of patients with gastric cancer based on the co-expression of B7H3 and HIF-1α. (E) Survival curve of patients with gastric cancer based on the co-expression of B7H3 and nuclear HIF-1α.

Article Snippet: Subsequently, the sections were incubated overnight at 4°C with mouse anti-human B7H3 antibody (R&D Systems, MN, USA, #AF1027, 1:200) or rabbit anti-human HIF-1α antibody(R&D Systems, MN, USA, #MAB19352,Clone #2443C,1:200).

Techniques: Expressing

Journal: Frontiers in Oncology

Article Title: Association of B7H3 with HIF-1α nuclear expression indicates poor prognosis and therapeutic potential in gastric cancer

doi: 10.3389/fonc.2026.1744341

Figure Lengend Snippet: (A) HIF-1α protein expression in HGC-27 cells under gradient concentrations of cobalt chloride induction. (B) B7H3 protein expression in HGC-27 cells under gradient concentrations of cobalt chloride (40 μmol/L, 200 μmol/L, 1000 μmol/L) induction. (C) HIF-1α protein expression in HGC-27 cells with overexpressed or knocked-down B7H3 under hypoxic and normal conditions. (D) HIF-1α expression in nuclear and cytoplasmic proteins of HGC-27 cells with overexpressed or knocked-down B7H3 under cobalt chloride induction. (E) Fluorescence immunohistochemistry of HGC-27 cells with B7H3 knockdown (SHB),B7H3 overexpression (LVB) and control group (SHN/LVN). Blue fluorescence represents DAPI, and red fluorescence represents HIF-1α. Scale bar: 10 μm.

Article Snippet: Subsequently, the sections were incubated overnight at 4°C with mouse anti-human B7H3 antibody (R&D Systems, MN, USA, #AF1027, 1:200) or rabbit anti-human HIF-1α antibody(R&D Systems, MN, USA, #MAB19352,Clone #2443C,1:200).

Techniques: Expressing, Fluorescence, Immunohistochemistry, Knockdown, Over Expression, Control

Journal: bioRxiv

Article Title: B7-H3-targeted natural killer cells effectively kill atypical teratoid / rhabdoid tumors and extend survival in orthotopic xenografts

doi: 10.64898/2026.01.15.699746

Figure Lengend Snippet: (A) Immunohistochemistry (IHC) staining for B7-H3 on FFPE primary tumor tissue, and (B) semi-quantitative scoring of B7-H3 expression. 20X magnification, n=8. (C) AT/RT cell line B7-H3 expression. Isotype-stained samples are shown in the histograms directly below each B7-H3-stained sample. (D) B7-H3 surface molecules per cell quantified by molecules of equivalent soluble fluorochrome (MESF) flow cytometry assay. Each dot representative of technical replicates. (E) Total B7-H3 protein expression with or without deglycosylation assessed by western blot.

Article Snippet: Briefly, 4 μm sections were stained with an anti-human B7-H3 primary antibody (clone D9M2L, Cell Signaling Technology 14058) on the Ventana Benchmark immunostaining system (Roche Diagnostics).

Techniques: Immunohistochemistry, Expressing, Staining, Flow Cytometry, Western Blot

Journal: bioRxiv

Article Title: B7-H3-targeted natural killer cells effectively kill atypical teratoid / rhabdoid tumors and extend survival in orthotopic xenografts

doi: 10.64898/2026.01.15.699746

Figure Lengend Snippet: (A) CARs consisting of Hu8H9 scFv and various affinity mutants, 2B4 (CD244) hinge (H), transmembrane (TM), and intracellular domains (IC), and CD3ζ signaling domain. (B) Representative primary NK-cell CAR expression. (C) CAR expression (n=4 healthy donors, 4-9 separate experiments). (D) Fold expansion of CAR-NK cells starting at day of transduction (n=4 healthy donors). (E) Representative CAR transduced NK-cell B7-H3 expression 2-4 days post-transduction. (F) B7-H3 expression on CAR-NK cells. (n = 3 healthy donors). *** = p<0.001 UTD vs. all others. (G) Representative measurement of CAR-NK cell degranulation post 2-hour co-culture with CHLA-06 AT/RT cells detected by CD107a staining.

Article Snippet: Briefly, 4 μm sections were stained with an anti-human B7-H3 primary antibody (clone D9M2L, Cell Signaling Technology 14058) on the Ventana Benchmark immunostaining system (Roche Diagnostics).

Techniques: Expressing, Transduction, Co-Culture Assay, Staining

Journal: bioRxiv

Article Title: B7-H3-targeted natural killer cells effectively kill atypical teratoid / rhabdoid tumors and extend survival in orthotopic xenografts

doi: 10.64898/2026.01.15.699746

Figure Lengend Snippet: (A) CHLA-06.ffLuc (25k cells) were injected into caudate putamen of NSG mice at day 0. Randomization/cohorting was performed at day 2, and intratumoral NK cell treatment (3 million cells per treatment) was started on day 3 and repeated on days 10 and 17. (B) Tumor radiance measured by IVIS Spectrum bioluminescent imaging and analyzed using Living Image software. (C) tumor radiance over time of CHLA-06.ffLuc bearing mice (n=8-9 per condition). (D) Kaplan-Meier survival curve. ***= p<0.001. (E) Brain tissue harvested at endpoint (D23 post-tumor, D6 post-third NK-cell treatment for both samples), representative slides from UTD NK treated and CAR-NK treated mice stained for human B7-H3 and CD45. 40X magnification.

Article Snippet: Briefly, 4 μm sections were stained with an anti-human B7-H3 primary antibody (clone D9M2L, Cell Signaling Technology 14058) on the Ventana Benchmark immunostaining system (Roche Diagnostics).

Techniques: Injection, Imaging, Software, Staining

Journal: bioRxiv

Article Title: B7-H3-targeted natural killer cells effectively kill atypical teratoid / rhabdoid tumors and extend survival in orthotopic xenografts

doi: 10.64898/2026.01.15.699746

Figure Lengend Snippet: (A) BT12.ffLuc (100k cells) were injected into the right lateral ventricle of NSG mice at day 0. Randomization/cohorting was performed at day 2, and ICV NK cell treatment was started on day 3 and repeated on days 10 and 17. (B) Kaplan-Meier survival curve, and (C) percent weight change over time of ICV BT12.ffLuc bearing mice. Each dotted line represents an individual animal. Representative histology of endpoint brain and spinal tissue from (D) UTD NK treated, and (E) CAR-NK treated mice with corresponding human B7-H3 and CD45 IHC stains. Animal endpoints = 31, 62 days post-tumor injection, respectively. 200X magnification. (F) Quantification of tumor associated human NK cells. n = 5-7 per condition.

Article Snippet: Briefly, 4 μm sections were stained with an anti-human B7-H3 primary antibody (clone D9M2L, Cell Signaling Technology 14058) on the Ventana Benchmark immunostaining system (Roche Diagnostics).

Techniques: Injection