anti cd81 pevio770  (Miltenyi Biotec)

Journal: bioRxiv

Article Title: Seminal extracellular vesicles from boar AI doses contain fertility-predictive protein and miRNA cargo and improve sperm physiology

doi: 10.64898/2026.03.16.712050

Figure Lengend Snippet: Cytometry plots and histograms of seminal extracellular vesicles (EVs) characterization. A) Flow cytometry beads of know size (200 nm – 2 µm). B) GFP EVs were used to identify EVs population. C) PBS 1X 0.22 μM filtered. D) Negative control of CellTrace Violet (CTV) staining diluted in PBS. E) High fertility (HF) and reduced fertility (RF) unstained EVs. F) HF CTV-stained EVs. G) RF CTV-stained EVs. H) CTV-stained EVs + human anti CD81-APC antibody (1:50). I) CTV-stained EVs + human anti CD63-PE antibody (1:50). J) CTV-stained EVs + human anti Hsp70-FITC antibody (1:50). K) Lysated EVs with Triton 1X + 0.1% SDS. Photodetectors used: Violet Side Scatter – Height (Violet SSC-H), Blue 525-Area (B525-A), Blue 525-Height (B525-H), Violet 450-Height (V450-H), Blue 585-Height (B585-H) and Red 660-Height (R660-H).

Article Snippet: To immunophenotype the EVs, 25 μL of samples (aproximately 2.0 x 10 9 EVs/mL) were incubated 30 min at 38 °C with conjugated antibodies against specific EVs markers: human anti CD63-PE diluted 1:50 (Ref. 130-118-150, Miltenyi Biotec, Germany), human anti CD81-APC 1:50 (Ref. 130-119-825, Miltenyi Biotec) and human anti Hsp70-FITC 1:50 (Ref. 130-124-700, Miltenyi Biotec).

Techniques: Cytometry, Flow Cytometry, Negative Control, Staining

Journal: BMC Psychiatry

Article Title: Normalization strategies for protein quantification in plasma astrocyte-derived extracellular vesicles: a clinical applicability study

doi: 10.1186/s12888-026-07796-6

Figure Lengend Snippet: Distribution of particle counts and EV biomarker levels in plasma ADEVs. ( A ) Distribution of ADEV particle counts measured by nano-flow cytometry (NFCM) and nanoparticle tracking analysis (NTA). ( B ) Distribution of five EV biomarkers (CD9, CD63, CD81, TSG101, and Alix) in ADEVs

Article Snippet: The levels of CD9 (Cat# CSB-EL004969HU, CUSABIO, China), CD63 (Cat# CSB-E14107h-IS, CUSABIO, China), CD81 (Cat# CSB-EL004960HU-IS, CUSABIO, China), Alix (Cat# CSB-EL017673HU, CUSABIO, China), TSG101 (Cat# CSB-EL025125HU, CUSABIO, China), and BDNF (Cat# E-EL-H0010, Elabscience, China) in ADEVs were measured using enzyme-linked immunosorbent assay (ELISA) kits.

Techniques: Biomarker Discovery, Clinical Proteomics, Flow Cytometry

Journal: BMC Psychiatry

Article Title: Normalization strategies for protein quantification in plasma astrocyte-derived extracellular vesicles: a clinical applicability study

doi: 10.1186/s12888-026-07796-6

Figure Lengend Snippet: Brain-derived neurotrophic factor (BDNF) Levels in astrocyte-derived extracellular vesicles (ADEVs) of MDD and HCs Under Various Normalization Strategies ( A – H ). ( I ) Receiver operating characteristic (ROC) curves of target proteins in ADEVs. The results showed that the area under the curve (AUC) for BDNF/CD9 was 0.738, with a sensitivity of 80.0% and a specificity of 66.7%, while the AUC for BDNF/CD81 was 0.756, with a sensitivity of 93.3% and a specificity of 53.3%. Note: n.s: not significant; * P < 0.05

Article Snippet: The levels of CD9 (Cat# CSB-EL004969HU, CUSABIO, China), CD63 (Cat# CSB-E14107h-IS, CUSABIO, China), CD81 (Cat# CSB-EL004960HU-IS, CUSABIO, China), Alix (Cat# CSB-EL017673HU, CUSABIO, China), TSG101 (Cat# CSB-EL025125HU, CUSABIO, China), and BDNF (Cat# E-EL-H0010, Elabscience, China) in ADEVs were measured using enzyme-linked immunosorbent assay (ELISA) kits.

Techniques: Derivative Assay

Journal: bioRxiv

Article Title: Ångström Resolution with Flow Immunofluorescence Localization Microscopy (FILM)

doi: 10.64898/2025.12.16.694546

Figure Lengend Snippet: (A) Due to the extremely high collection efficiency of FILM, out-of-focus events are also robustly detected, generating images of a central peak surrounded by multiple concentric diffractive rings. Simple threshold detection over the small red detection windows (9×9 pixels) will therefore pick up the same particle multiple times, as observed for several out-of-focus events in the displayed 5 s frame taken from the CD81 staining of healthy tonsil tissue . For most datasets, it was therefore necessary to filter the thresholded events for radial symmetry by determining the center of radial symmetry for each event within the detection window (see Materials and Methods) and comparing it with the center of its 2D Gaussian fit. (B) Removal of events with more than 1-pixel (103.17 nm) deviation of their radial and Gaussian centers, followed by probe tracking over the entire acquisition, resulted in the displayed, color-coded, tracked events for the frame shown in A. Note the absence here of most of the spurious detections on the ring-like structures in A.

Article Snippet: α-tubulin (REA1136 REAfinityTM, h, APC, #130-119-541, Miltenyi Biotec), CD2 (REA972 REAfinityTM, h, APC, #130-116-150, Miltenyi Biotec), CD20 (REA780 REAfinityTM, h, APC, #130-111-339, Miltenyi Biotec), CD28 (REA612 REAfinityTM, h, APC, #130-118-343, Miltenyi Biotec), CD45 (REA1023 REAfinityTM, h/nhp, APC, #130-117-191, Miltenyi Biotec), CD54/ICAM-1 (REA266 REAfinityTM, h, APC, #130-120-711, Miltenyi Biotec), CD81 (REA513 REAfinityTM, h, APC, #130-119-787, Miltenyi Biotec), CD107a/LAMP1 (REA792 REAfinityTM, h, APC, #130-111-847, Miltenyi Biotec), IgD (REA740 REAfinityTM, h, APC, #130-110-644, Miltenyi Biotec), IgM (REAL689 REAdye_leaseTM, h, PE, #130-125-786, Miltenyi Biotec), Pan-Cytokeratin (REA1141 REAfinityTM, h, APC, #130-120-096, Miltenyi Biotec), β-catenin (REA480 REAfinityTM, h, APC, #130-124-444, Miltenyi Biotec), TCRα/β (REA652 REAfinityTM, h, APC, #130-113-535, Miltenyi Biotec), TOM22 (REA1185 REAfinityTM, APC, h/ms, #130-122-079, Miltenyi Biotec), DAPI Staining Solution (#130-111-570, Miltenyi Biotec), and ATTO 643 phalloidin (ATT-AD643-81, ATTO-TEC).

Techniques: Staining

Journal: bioRxiv

Article Title: Ångström Resolution with Flow Immunofluorescence Localization Microscopy (FILM)

doi: 10.64898/2025.12.16.694546

Figure Lengend Snippet: (A–C) FILM acquisition of antibodies labeled with APC that recognize TCRα/β, CD28, and CD81 in the cSMAC (10 nm pixels with 3-pixel Gaussian blurring). Cyan boxed region shows the IF image (103.17 nm pixels) for the corresponding white boxed region. (D–F) Zoom-in of the respective boxed regions for the panels to the left. Localizations are shown as peak-normalized 2D Gaussians with color-coded precision according to the colorbar. (G–I) Zoom-in of the respective boxed regions for the panels to the immediate left. Localizations are shown as crosses (indicating ±1-σ uncertainty in x and y ) with color-coded precision according to the colorbar.

Article Snippet: α-tubulin (REA1136 REAfinityTM, h, APC, #130-119-541, Miltenyi Biotec), CD2 (REA972 REAfinityTM, h, APC, #130-116-150, Miltenyi Biotec), CD20 (REA780 REAfinityTM, h, APC, #130-111-339, Miltenyi Biotec), CD28 (REA612 REAfinityTM, h, APC, #130-118-343, Miltenyi Biotec), CD45 (REA1023 REAfinityTM, h/nhp, APC, #130-117-191, Miltenyi Biotec), CD54/ICAM-1 (REA266 REAfinityTM, h, APC, #130-120-711, Miltenyi Biotec), CD81 (REA513 REAfinityTM, h, APC, #130-119-787, Miltenyi Biotec), CD107a/LAMP1 (REA792 REAfinityTM, h, APC, #130-111-847, Miltenyi Biotec), IgD (REA740 REAfinityTM, h, APC, #130-110-644, Miltenyi Biotec), IgM (REAL689 REAdye_leaseTM, h, PE, #130-125-786, Miltenyi Biotec), Pan-Cytokeratin (REA1141 REAfinityTM, h, APC, #130-120-096, Miltenyi Biotec), β-catenin (REA480 REAfinityTM, h, APC, #130-124-444, Miltenyi Biotec), TCRα/β (REA652 REAfinityTM, h, APC, #130-113-535, Miltenyi Biotec), TOM22 (REA1185 REAfinityTM, APC, h/ms, #130-122-079, Miltenyi Biotec), DAPI Staining Solution (#130-111-570, Miltenyi Biotec), and ATTO 643 phalloidin (ATT-AD643-81, ATTO-TEC).

Techniques: Labeling

Journal: bioRxiv

Article Title: Ångström Resolution with Flow Immunofluorescence Localization Microscopy (FILM)

doi: 10.64898/2025.12.16.694546

Figure Lengend Snippet: (A) Overview DAPI image (using a 10× objective) of a PFA-fixed and quenched human palatine tonsil tissue section (4 µm thick) containing a follicle. White borders (dotted lines) are used to indicate the anatomic follicular zones corresponding to the germinal center (GC), mantle zone (MZ) and interfollicular region (IFR). (B) Multicolor overlay of sequential FILM acquisitions of the boxed region in A of PE-labeled IgM at 20% LED power (0.7 W cm −2 ) and APC-labeled IgD, CD81, and CD20 at 100% LED power (5.3 W cm −2 ) (10-nm pixels with 3-pixel Gaussian blurring). The sample was washed extensively with PBS under continuous flow and photobleaching before application of each successive probe. After FILM imaging, IF images were acquired (shown on the left side of the panel for comparison). (C) Zoom-in of the boxed region with Ć label in B (10-nm pixels with 3-pixel Gaussian blurring). (D) Zoom-in of boxed region in C using peak-normalized Gaussian representations for the localizations (color-coded for each marker as in B). (E) Zoom-in of boxed region in D. Colored arrows indicate individual localizations with 2.1 nm and 1.3 nm precision for IgD and CD20, respectively. (F) Zoom-in of the boxed region with F’ label in B (10-nm pixels with 3-pixel Gaussian blurring) overlaid with a conventional DAPI image (grayscale). Boxed region contains a single isolated linear CD81 structure. (G) Zoom-in of the boxed region in F, now displaying only the CD81 localizations as peak-normalized Gaussians with color-coded precision according to the colorbar. The estimated positions (centers of the Gaussians) for all 27 individual localizations lie within the overlaid strip of width 40 nm (two offset localizations toward the upper right were not considered as part of the structure). See for four additional examples of similar linear CD81 structures.

Article Snippet: α-tubulin (REA1136 REAfinityTM, h, APC, #130-119-541, Miltenyi Biotec), CD2 (REA972 REAfinityTM, h, APC, #130-116-150, Miltenyi Biotec), CD20 (REA780 REAfinityTM, h, APC, #130-111-339, Miltenyi Biotec), CD28 (REA612 REAfinityTM, h, APC, #130-118-343, Miltenyi Biotec), CD45 (REA1023 REAfinityTM, h/nhp, APC, #130-117-191, Miltenyi Biotec), CD54/ICAM-1 (REA266 REAfinityTM, h, APC, #130-120-711, Miltenyi Biotec), CD81 (REA513 REAfinityTM, h, APC, #130-119-787, Miltenyi Biotec), CD107a/LAMP1 (REA792 REAfinityTM, h, APC, #130-111-847, Miltenyi Biotec), IgD (REA740 REAfinityTM, h, APC, #130-110-644, Miltenyi Biotec), IgM (REAL689 REAdye_leaseTM, h, PE, #130-125-786, Miltenyi Biotec), Pan-Cytokeratin (REA1141 REAfinityTM, h, APC, #130-120-096, Miltenyi Biotec), β-catenin (REA480 REAfinityTM, h, APC, #130-124-444, Miltenyi Biotec), TCRα/β (REA652 REAfinityTM, h, APC, #130-113-535, Miltenyi Biotec), TOM22 (REA1185 REAfinityTM, APC, h/ms, #130-122-079, Miltenyi Biotec), DAPI Staining Solution (#130-111-570, Miltenyi Biotec), and ATTO 643 phalloidin (ATT-AD643-81, ATTO-TEC).

Techniques: Labeling, Imaging, Comparison, Marker, Isolation, Stripping Membranes

Journal: bioRxiv

Article Title: Ångström Resolution with Flow Immunofluorescence Localization Microscopy (FILM)

doi: 10.64898/2025.12.16.694546

Figure Lengend Snippet: (A–D) On the left are further examples of linear distributions of CD81 (see ) that extend out into regions of the tissue with significantly lower nuclear staining. On the right are zoom-ins of the boxed regions indicated in the respective left panels. These depict only the CD81 localizations as peak-normalized 2D Gaussians with color-coded precision given by the colorbar. Almost all of the localizations (centers of the Gaussians) lie within the overlaid 50 nm reference strips. Localizations with significant offsets from the strip were consistent with the otherwise diffuse CD81 staining.

Article Snippet: α-tubulin (REA1136 REAfinityTM, h, APC, #130-119-541, Miltenyi Biotec), CD2 (REA972 REAfinityTM, h, APC, #130-116-150, Miltenyi Biotec), CD20 (REA780 REAfinityTM, h, APC, #130-111-339, Miltenyi Biotec), CD28 (REA612 REAfinityTM, h, APC, #130-118-343, Miltenyi Biotec), CD45 (REA1023 REAfinityTM, h/nhp, APC, #130-117-191, Miltenyi Biotec), CD54/ICAM-1 (REA266 REAfinityTM, h, APC, #130-120-711, Miltenyi Biotec), CD81 (REA513 REAfinityTM, h, APC, #130-119-787, Miltenyi Biotec), CD107a/LAMP1 (REA792 REAfinityTM, h, APC, #130-111-847, Miltenyi Biotec), IgD (REA740 REAfinityTM, h, APC, #130-110-644, Miltenyi Biotec), IgM (REAL689 REAdye_leaseTM, h, PE, #130-125-786, Miltenyi Biotec), Pan-Cytokeratin (REA1141 REAfinityTM, h, APC, #130-120-096, Miltenyi Biotec), β-catenin (REA480 REAfinityTM, h, APC, #130-124-444, Miltenyi Biotec), TCRα/β (REA652 REAfinityTM, h, APC, #130-113-535, Miltenyi Biotec), TOM22 (REA1185 REAfinityTM, APC, h/ms, #130-122-079, Miltenyi Biotec), DAPI Staining Solution (#130-111-570, Miltenyi Biotec), and ATTO 643 phalloidin (ATT-AD643-81, ATTO-TEC).

Techniques: Staining, Stripping Membranes

Journal: bioRxiv

Article Title: Ångström Resolution with Flow Immunofluorescence Localization Microscopy (FILM)

doi: 10.64898/2025.12.16.694546

Figure Lengend Snippet: (A) Representative fluorescence image (5 s exposure; 103.17 nm pixels) from the CD81 dataset (see ) showing simultaneous detection of both in-focus and out-of-focus antibody probes conjugated to a single APC. (B–G) Zoom-in of the boxed region in A for the six consecutive frames over which the out-of-focus probe appeared (30 s lifetime). Note the appearance and disappearance of other in-focus localizations. (H) Summed image of all frames from panels B–G, with five diffractive rings clearly discernible. In-focus localizations are indicated that do not belong to the PSF of the out-of-focus probe. Roughly 5.4 million photons were detected for this out-of-focus probe. (I) A Gibson-Lanni model for the PSF of a probe located 4.6 µm below the focal plane (with focal plane positioned at 5 µm above the coverslip glass) exhibited a highly similar pattern as that found for the probe in H. The model was calculated using the plugin PSFgenerator , with values for the positions of the focal plane and probe chosen empirically to give a good match to H. Further values required for the model were the coverslip thickness (170 µm) and the refractive indices of the oil immersion (1.518), borosilicate coverslip glass (1.53) and water-based buffer (1.33).

Article Snippet: α-tubulin (REA1136 REAfinityTM, h, APC, #130-119-541, Miltenyi Biotec), CD2 (REA972 REAfinityTM, h, APC, #130-116-150, Miltenyi Biotec), CD20 (REA780 REAfinityTM, h, APC, #130-111-339, Miltenyi Biotec), CD28 (REA612 REAfinityTM, h, APC, #130-118-343, Miltenyi Biotec), CD45 (REA1023 REAfinityTM, h/nhp, APC, #130-117-191, Miltenyi Biotec), CD54/ICAM-1 (REA266 REAfinityTM, h, APC, #130-120-711, Miltenyi Biotec), CD81 (REA513 REAfinityTM, h, APC, #130-119-787, Miltenyi Biotec), CD107a/LAMP1 (REA792 REAfinityTM, h, APC, #130-111-847, Miltenyi Biotec), IgD (REA740 REAfinityTM, h, APC, #130-110-644, Miltenyi Biotec), IgM (REAL689 REAdye_leaseTM, h, PE, #130-125-786, Miltenyi Biotec), Pan-Cytokeratin (REA1141 REAfinityTM, h, APC, #130-120-096, Miltenyi Biotec), β-catenin (REA480 REAfinityTM, h, APC, #130-124-444, Miltenyi Biotec), TCRα/β (REA652 REAfinityTM, h, APC, #130-113-535, Miltenyi Biotec), TOM22 (REA1185 REAfinityTM, APC, h/ms, #130-122-079, Miltenyi Biotec), DAPI Staining Solution (#130-111-570, Miltenyi Biotec), and ATTO 643 phalloidin (ATT-AD643-81, ATTO-TEC).

Techniques: Fluorescence