fluorescein labeled lectin  (Vector Laboratories)

Journal: Science Advances

Article Title: A conserved glycan motif induces broadly reactive functional antibodies against the zoonotic pathogen Streptococcus suis

doi: 10.1126/sciadv.adz1854

Figure Lengend Snippet: ( A ) Binding of plant lectins to surface structures of capsule-deficient S. suis strains (ΔCPS). SBA binds to N -acetylgalactosamine (GalNAc) and, to a lesser extent, galactose (Gal); RCA 120 binds to both Gal and GalNAc; sWGA has a special affinity to N -acetylglucosamine (GlcNAc). Data from biological triplicates are presented as mean values ± SD. ( B ) Glycosyl composition analysis by GC-MS of TMS (trimethylsilyl) derivatives of methyl glycosides of S. suis RPS from S10 and 861160 released by mild acid hydrolysis after chemical N -acetylation. ( C ) Plant lectin binding to isolated RPS from S. suis S10 and 861160. Data show technical triplicates (mean values ± SD) and are representative for two independent experiments. ( D and E ) Presence and absence of the most abundant glycosyl linkage residues (D) and phosphate (E) of S. suis RPS from S10 and 861160. Glycosyl linkage residues were analyzed by GC-MS of partially methylated alditol acetate derivatives. Phosphate was determined by malachite green assay following hydrolysis with hydrochloric acid and digestion with alkaline phosphatase. Original data is in tables S2 and S3.

Article Snippet: Subsequently, 12.5 μl of bacterial suspension was incubated in a 96-well V-bottom plate with the same volume of pig serum or fluorescein-labeled lectin (Vector Laboratories, FLK-2100 and FL-1021S-5).

Techniques: Binding Assay, Gas Chromatography-Mass Spectrometry, Isolation, Methylation, Malachite Green Assay

Journal: Nature Communications

Article Title: ARMH3 is an ARL5 effector that promotes PI4KB-catalyzed PI4P synthesis at the trans -Golgi network

doi: 10.1038/s41467-024-54410-y

Figure Lengend Snippet: a WT and KO HeLa cells were immunostained for endogenous GOLPH3. Images are shown in grayscale with nuclei (DAPI) in blue. Scale bars: 10 μm. Notice the decrease in GOLPH3 staining in the KO cells. b Quantification of GOLPH3 intensity at the Golgi complex relative to the cytosol from experiments such as that shown in panel a. Data are represented as SuperPlots showing the mean ± SD of the means of the individual data points from three independent, color-coded experiments. The statistical significance of the differences was calculated by one-way ANOVA with multiple comparisons using Dunnett’s test. c Lectin blotting of WT and KO HeLa cells with HPA-Alexa Fluor 647 and PNA-Fluorescein. Immunoblotting for α-tubulin was used as a loading control. Arrows indicate glycoprotein species whose intensity is changed in the KO cells. Blots are representative of two independent experiments. d Immunoblot analysis of WT, KO, and rescue (ARL5B-mCherry, HA-ARMH3, or Myc-PI4KB stably expressed in corresponding KO cell lines) HeLa cells using antibodies to LAMP1 and α-tubulin (control). Similar results were obtained from two independent biological replicates. e WT and KO HeLa cells, untreated or treated with PNGase F, were analyzed by immunoblotting with an antibody to LAMP1. The positions of molecular mass markers (kDa) in panels c – e are indicated on the left. Blots are representative of three independent experiments. f Schematic representation of the role of the SYS1-ARFRP1-ARL5-ARMH3-PI4KB axis in PI4P synthesis at the TGN for recruitment of GOLPH3 to the TGN. Created in BioRender. Bonifacino, J. (2024) BioRender.com/o49h844. Source data are provided in the Source Data file.

Article Snippet: Lectin Helix pomatia (HPA)-Alexa Fluor 647 (Invitrogen, L32454) and Arachis Hypogaea (peanut) Agglutinin (PNA)-Fluorescein from Lectin Kit I, Fluorescein (Vector Laboratories, FLK-2100) were used for the lectin blot assays.

Techniques: Staining, Western Blot, Control, Stable Transfection

Journal: Scientific Reports

Article Title: Probing the expression and adhesion of glycans involved in Helicobacter pylori infection

doi: 10.1038/s41598-024-59234-w

Figure Lengend Snippet: Lectin binding specificities.

Article Snippet: The lectins used for immunofluorescence were Fluorescein Lectin Kit I (FLK-2100) and Fluorescein Lectin Kit II (FLK-3100) supplied by Vector laboratories.

Techniques: Binding Assay

Journal: Scientific Reports

Article Title: Probing the expression and adhesion of glycans involved in Helicobacter pylori infection

doi: 10.1038/s41598-024-59234-w

Figure Lengend Snippet: Lectin binding to H. pylori laboratory strains. ( A ) ELISA screening of the Lectin Array against H. pylori SS1 lysate ( B ) ELISA screening of the Lectin Array against H. pylori 26695 lysate. Pool A is a technical control (pooled H. pylori SS1 specific mouse serum). * p ≤ 0.05 by one-way ANOVA vs negative control. Each sample was run in quadruplicate in three separate assays, error bars represent standard deviation.

Article Snippet: The lectins used for immunofluorescence were Fluorescein Lectin Kit I (FLK-2100) and Fluorescein Lectin Kit II (FLK-3100) supplied by Vector laboratories.

Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay, Control, Negative Control, Standard Deviation

Journal: Scientific Reports

Article Title: Probing the expression and adhesion of glycans involved in Helicobacter pylori infection

doi: 10.1038/s41598-024-59234-w

Figure Lengend Snippet: Lectin binding to surface of H. pylori laboratory strains. ( A ) Immunofluorescence images of lectins against H. pylori SS1. Con A WGA and sWGA appear to bind strongly ( B ) Immunofluorescence images of lectins against H. pylori 26695. Con A appears to bind moderately, while WGA and sWGA show a stronger binding in IF. The lectins PA, RCA, SBA, UEA, DBA, SJA, PVL, PSA, GSC, LCA. PHA-E, and GSL were also tested in immunofluorescence and showed no reactivity (Fig. S1). The white scale bar represents 2 μm.

Article Snippet: The lectins used for immunofluorescence were Fluorescein Lectin Kit I (FLK-2100) and Fluorescein Lectin Kit II (FLK-3100) supplied by Vector laboratories.

Techniques: Binding Assay, Immunofluorescence