Human CD39L3/ENTPD3 Antibody

Detection of CD39L3/ENTPD3 by Immunocytochemistry/ Immunofluorescence

ImmTAAI binding and labeling of beta cells in pancreas slices is concentration dependent.(A) Dark-field stereomicroscopy image of a pancreas slice showing islets outlined in red. (B) Pancreas slice incubated with CF647-labeled PPI ImmTAAI and subsequently fixed and stained for insulin, glucagon, and ENTPD3, and imaged by confocal. (C) Confocal microscopy images of live pancreas slices incubated with CF647-labeled PPI ImmTAAI at 2, 20, or 200 nM for 1 hour and stained with a labeled primary antibody for cell surface ENTPD3. The off-target CF647-labeled Tel ImmTAAI was used as a control. (D) MFI quantification of confocal microscopy images showing the dose-dependent binding of PPI ImmTAAI to tissue slices. (E) Pearson’s coefficient colocalization analysis of ImmTAAI fluorescent signal with anti-ENTPD3 fluorescent signal. PPI ImmTAAI colocalizes with anti-ENTPD3 well at all three concentrations tested, indicating that beta cell–specific binding is maintained in native tissue. Individual dots represent measurements per islet (means from one donor for Tel ImmTAAI and two donors for PPI ImmTAAI labeled). Statistical differences were determined by two-way analysis of variance (ANOVA) followed by Tukey’s post hoc analysis. ns, not significant; ***P < 0.001, ****P < 0.0001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/41920992), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of CD39L3/ENTPD3 by Immunocytochemistry/ Immunofluorescence

ImmTAAI binding is HLA specific in tissue slices.(A) Confocal microscopy images of pancreas slices incubated with 20 nM CF647-labeled PPI ImmTAAI for 2 hours (h) and costained for ENTPD3. No ImmTAAI staining is observed in slices from HLA-A*31 tissue. Islet granularity (reflectance) and viability (SYTOX blue) are also shown. (B) MFI analysis of ImmTAAI binding in HLA-A2–negative and –positive tissue, showing binding only in HLA-A2–positive tissue. (C) Pearson’s coefficient colocalization analysis of ImmTAAI fluorescent signal with anti-ENTPD3 fluorescent signal. ImmTAAI signal colocalizes with ENTPD3 only in HLA-A2–positive tissue. Each dot represents a separate islet from one or two separate slices per condition (n = 1 donor for Tel ImmTAAI in HLA-A*31 tissue, n = 2 donors for HLA-A2) and is the mean of three to six z-planes taken per islet. (D) Confocal microscopy images of pancreas slices treated with a 5-hour pulse of IFN-gamma (100 IU/ml) and then fixed after 24 hours and stained for insulin and HLA-ABC. (E) MFI analysis of HLA-ABC expression in islet and exocrine regions. Each dot represents a separate islet/exocrine region image from ≥three slices per condition. (F) Live confocal microscopy images of pancreas slices treated with a 5-hour pulse of IFN-gamma (100 IU/ml) and incubated with 20 nM CF647-labeled PPI ImmTAAI at 2, 6, or 18 hours. (G) MFI analysis of ImmTAAI signal at each time point post–IFN-gamma treatment. Each dot represents a separate islet from ≥three slices per condition. NT, no treatment. Statistical differences were determined by two-way ANOVA followed by Tukey’s post hoc analysis. ns, not significant. **P < 0.01, ***P < 0.001, ****P < 0.0001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/41920992), licensed under a CC-BY license. Not internally tested by R&D Systems.