b4 ib4 fl 1201 fluorescein  (Vector Laboratories)

Journal: bioRxiv

Article Title: A cell membrane-hybridized nanocarrier-based RNA delivery system for ocular neovascular diseases

doi: 10.64898/2025.12.23.696136

Figure Lengend Snippet: (a) Fluorescence staining images showing the distribution of liposomes in the choroidal flat-mounts 7 days after intravitreal injection, with neovascular lesions (IB4, green) and liposomes (Cy3, red). (b) Statistical results of fluorescence intensity for each group.

Article Snippet: Griffonia Simplicifolia Lectin I isolectin B4 (IB4) (FL-1201, Fluorescein) was purchased from vector laboratories (USA).

Techniques: Fluorescence, Staining, Liposomes, Injection

Journal: Frontiers in Physiology

Article Title: Tmem45b modulates itch via endoplasmic reticulum calcium regulation

doi: 10.3389/fphys.2025.1708686

Figure Lengend Snippet: Tmem45b is primarily expressed in itch-sensing neurons. (A) Immunostaining results show the expression of Tmem45b (green), CGRP (red), and NF200 (red). (B) Immunostaining results show the co-expression of Tmem45b (green) and IB4 (red). (C) Statistical analysis shows the proportion of Tmem45b + IB4 + . (D) scRNA-seq (10x Genomics) analysis identified 16 subtypes of DRG neurons. Gene annotations for these 16 neuron types are shown in the right panel. Tmem45b is predominantly enriched in neurons positive for Mrgprd, Mrgpra3, Nppb , and Th . (E) RNAscope in situ hybridization shows co-localization among Tmem45b (green), Mrgprd (red) , Mrgpra3 (red), and Nppb (red). Immunostaining results show that Tmem45b (green) is co-expressed with Th (red). Arrows indicate co-expressing cells. Scale bar, 20 μm. (F) Proportions of Mrgprd + , Mrgpra3 + , Nppb + , and Th + neurons among Tmem45b + DRG neurons. DRG was obtained from at least 3 mice.

Article Snippet: Antibodies used in this study: GM130 (BD, 610822), Th (Millipore, AB1542), IB4 (Vector, FL-1201-.5), Tuj1 (Starter, SDT-251-28), CGRP (Dia Sorin, 24112), NF200 (CST, 2836S), PDI (Santa Cruz, SC-20132), Calnexin (Abcam, ab112995), Serca1 (Proteintech, 22361-1-AP), mitochondria-tracer (Beyotime, C1048), TGN38 (Bio-Rad, AHP499G), GFAP (Millipore, MAB3402), IBA1 (Abcam, ab5076).

Techniques: Immunostaining, Expressing, RNAscope, In Situ Hybridization

Journal: Regenerative Therapy

Article Title: Assessment of permeability in deep tissue capillaries using a new method reflects the nutrient supply status in a healthy heart

doi: 10.1016/j.reth.2025.10.005

Figure Lengend Snippet: Detection of fluorescently labeled dextran leaking from cardiac capillaries A. Three fractionations by fluorescence intensity: background (BG), extra-capillary (BG-BG x a), intra-capillary (>BG x a). B. Methods for calculating background. FITC-conjugated GSL-IB4 was injected into the tail vein 5 min after PBS injection, and another 5 min later, mice were sacrificed, the cardiac tissue was removed, and promptly frozen with isopentane onto dry ice. Frozen tissue sections were cut transversely at 10 μm using the cryostat, and the sections were mounted onto slides. The slides were dried thoroughly and observed under a fluorescence microscope. Twenty 150 μm × 150 μm per 1 section were cut out from the left ventricular wall, where capillaries were observed parallel to the myocardial layer, using LAS X software (Leica), and saved as TIFF (922 x 922 pixels; 22461.544 μm 2 /1 area). Twenty areas per section, three sections per mouse, and three (young) or two (aged) female mice were analyzed, resulting in a total of 180 or 120 images ( see Materials and Methods section for details ). Scale bar: 1 mm. C. Methods for calculating the amount of fluorescently labeled dextran leaked from capillaries. FITC-conjugated GSL-IB4 was injected into the tail vein 5 min after TD 40 injection, and another 5 min later, mice were sacrificed, the cardiac tissue was removed, and promptly frozen with isopentane onto dry ice. Frozen tissue cut sections were prepared in the same manner as above. Fluorescence images of dextran and GSL-IB4 are imported into ImageJ separately, converted to grayscale, and binarized by setting the threshold value for each. The ratio of fluorescence-detected areas to total area was calculated from the acquired fluorescent images. Three sections per mouse, with three mice per month, were analyzed for a total of 180 images. Scale bar: 50 μm.

Article Snippet: To visualize cardiac endothelial cells, fluorescein isothiocyanate (FITC) -conjugated GSL-IB4 (#FL-1201; Vector Laboratories Inc., Burlingame, CA, USA) was injected into the tail vein 5 min after TD 40 injection under anesthesia (1 mg/ml, 50 μl/10 g).

Techniques: Labeling, Fluorescence, Injection, Microscopy, Software

Journal: Genome Research

Article Title: Homozygous editing of multiple genes for accelerated generation of xenotransplantation pigs

doi: 10.1101/gr.279709.124

Figure Lengend Snippet: Selection of homozygously edited cells based on highest surrogate reporter expression. ( A ) Schematic of selection strategy using version 2.1- eGFP (V2.1G), a nonintegrated surrogate reporter system. PFFs were cotransfected with V2.1G, Cas9, and sgRNA plasmid, whereas the control group was only transfected with the V2.1G plasmid. FACS analysis revealed varying intensities of FITC expression among the cells. (H) The highest FITC intensity population, (M) the middle FITC intensity population, (L) the lower FITC intensity population, and (N) the negative FITC intensity population. ( B ) Correlation between genome editing efficiency and FITC intensity of endogenous targeting in PFFs. ( C ) Representative Sanger sequences of CMAH target site in wild-type cells, homozygous cell clones, and cell pools selected by V2.1G. ( D ) FACS analysis of GGTA1 targeting in PFF pools detection by IB4 lectin staining. ( E ) Western blot analysis of USE1-targeted Vero E6 cell pools with middle and high FITC intensity, respectively.

Article Snippet: The following lectins and antibodies were used for staining: IB4 lectin (Vector FL-1201, 1:50 dilution), DBA lectin (Vector FL-1031, 1:50 dilution), AntiNeuGC (BioLegend 146901, 1:50 dilution), and thrombomodulin antibody (Abmart T55279, 1:100 dilution).

Techniques: Selection, Expressing, Plasmid Preparation, Control, Transfection, Clone Assay, Staining, Western Blot