Thick (40 m) paraffin sections were stained with Wheat Germ Agglutinin-Alexa Fluor 488 to visualise cell membranes, and DAPI (4,6-diamidino-2-phenylindole) to visualise WGA is a lectin that specifically binds chitin [ 15 ], the The blocks were dyed with hematoxylin-eosin (HE), Sirius Red and Massons trichrome. Hi Michael. Thank you for your input! So far we have not been using HEPES buffer for the WGA staining. Can I ask you at which steps you use the HEP Staining with wheat germ agglutinin (leads to N-acetylglucosamine) occurs predominantly to each of the anuclear retinal layers and progressively disappears from each of the nuclear Thank you for your answer, Ute! I used fluorophore labelled WGA and I will now try to increase the concentration. Kind regards, Edward Dietary lectins are carbohydrate-binding proteins found in food sources. Wheat germ agglutinin. WGA is found abundantlybut not exclusivelyin the wheat kernel, where it got the 'germ' name from. In mammals the N-acetyl-D-glucosamine that WGA binds to is found in cartilage and cornea among other places. In those animals sialic acid is found in mucous membranes, e.g. the lining of the inner nose, and digestive tract. The use of wheat germ agglutinin (WGA) conjugated with a fluorophore is a very reliable alternative with a unique efficiency in resolving the finest details of fungal morphology . IHC electron microscopy (SEM) (B) and wheat germ agglutinin (WGA) staining (C). See the attached link. Description Wheat germ agglutinin (WGA) is one of the most widely used lectins in cell biology. Our Alexa Fluor 488 conjugate of WGA exhibits the bright, green fluorescence of the Alexa Fluor 488 dye (excitation/emission maxima 495/519 nm). Alexa Fluor 488 WGA binds to sialic acid and N -acetylglucosaminyl residues. (PBS) was used to stain tissue sections. We therefore used FITC-labeled wheat germ agglutinin (WGA), which marks fibrotic tissue in comparable quality as the established picrosirius red (SR) staining, for the staining of post myocardial infarction scar tissue. The fibrosis amount was quantified in a histogram-based approach using the non-commercial image processing program ImageJ. HI Edward. We've been using WGA on fixed tissue culture cells. I will say that one sticking point for us was the use of HEPES buffer. Without it, w Description SDS Pricing; 61767: lyophilized, 55% (GE), powder: Expand. The protein dissociates into monomers in acidic media, pK of the reaction 4. Contents and storage information. We No difference in staining was found using other lectins (Canavalia ensiformis agglutinin, Ricinus communis agglutinin, Lens culinaris agglutinin). All Photos (1) Lectin, immobilized on Agarose CL-4B from Triticum vulgaris. I would assume I've been trying to visualize the cell membrane in mouse heart sections, but so far I Paraffin sections prepared from prenatal rat brain (embryonic days 14, 16, 18 and 20) were incubated with buffered wheat germ agglutinin labelled with fluorescein isothiocyanate (FITC Irwin J. Goldstein Ph.D., Ronald D. Poretz Ph.D., in New Comprehensive Biochemistry, 1997 3.3.1 Wheat germ agglutinin. Match Criteria: Product Name, Keyword. Wheat germ agglutinin (WGA) contains a group of closely related isolectins, with an isoelectric point about pH 9. The use of established fibrosis staining techniques is limited on frozen cardiac tissue sections due to a reduced color contrast compared to paraffin embedded sections. Wheat germ agglutinin (WGA), even at low doses, demonstrated maximum toxicity toward acute myeloid leukemia (AML) cells. Technical Information. Helmut Plattner, in International Review of Cell and Molecular Biology, 2010. Wheat germ agglutinin (WGA) is a carbohydrate-binding protein of approx. Thick (40 m) paraffin sections were stained with Wheat Germ Agglutinin-Alexa Fluor 488 to visualise cell membranes, and DAPI (4,6-diamidino-2-phenylindole) to visualise Edward. According to the manual from Thermo, the WGA should be diluted in HBSS for staining. Afterwards, you can stain with additional antibodies. Hello Ashraf, we used the fluorophore labelled WGA because it was alreadyavailable in our lab at that moment. Do you have better experience with A for staining of tissue sections, or for western blotting. The use of established fibrosis staining techniques is limited on frozen cardiac Summary. As a consequence, I tried Molecular Probes fluorescently labeled wheat germ agglutinin. Histological staining. Edward. The Wheat Germ Agglutinin Sampler Kit includes introductory samples of four fluorescent wheat germ agglutinin conjugates (Alexa Fluor 350, Oregon Green 488, tetramethylrhodamine, and Texas RedX conjugates). Wheat germ agglutinin (WGA) is a lectin that protects wheat from insects, yeast and bacteria.An agglutinin protein, it binds to N-acetyl-D-glucosamine and Sialic acid. Paraffin sections were prepared by agar preembedding myocardial tissue. We Using AML cell lines, we show time- and dose-dependent killing by WGA. We therefore used *p0.05. Has anybody experience with WGA staining in formalin-fixed and paraffin-embedded tissue sections? The use of established fibrosis staining techniques is limited on frozen cardiac tissue sections due to a reduced color contrast compared to paraffin embedded sections. The use of established fibrosis staining techniques is limited on frozen cardiac tissue sections due to a reduced color contrast compared to paraffin embedded sections. The lectin is a homodimer composed of subunits of M r = 23 600 Da which dissociates into monomers at acid pH.. Each protomeric Wheat germ agglutinin (WGA), which binds to glycoproteins of the cell membrane, is routinely used for the staining of skeletal and cardiac sarcolemma to determine cross sectional area or myocyte density. The available dyes I tried never produced a good stain and I was not willing to go to the effort of making liposomes to deliver dye. 2020;2146:53 in AM root sections of Lotus japonicus. The use of established fibrosis staining techniques is limited on frozen cardiac tissue sections due to a reduced color contrast compared to paraffin embedded sections. Wheat germ agglutinin is the most studied of these lectins and also one of the most useful in its biomedical application. Wheat germ agglutinin selectively binds to N-acetylglucosaminyl (GlcNAc) and N-acetylneuraminic acid (sialic acid) residues on glycoconjugates and oligosaccharides found throughout the cell surface.The presence and distribution of these residues can not only vary among cell and organelle types but can also be expressed abnormally in various pathologies, The amino acid sequence provides a figure of 21,600, based on 171 amino acids per polypeptide chain. Wheat germ agglutinin staining as a suitable method for detection and quantification of fibrosis in cardiac tissue after myocardial infarction. 1. Heinrich-Heine University Dsseldorf. The quantification of fibrotic tissue is an important task in the analysis of cardiac remodeling. Immunohistochemistry stain (IHC)Paraffin . Compare Product No. (b) With Alcian blue staining (pH 2.5), a positive staining (dark blue) of the gland types is noted (white arrowheads). This lectin is used for the purification of insulin receptors. (D) Quantification of bone resorption area per view area in B. 4.2.2 Early and late endosomes. We also show Fluorescent Staining of Arbuscular Mycorrhizal Structures Using Wheat Germ Agglutinin (WGA) and Propidium Iodide Methods Mol Biol. Data is expressed as mean sd (n=15). Synonym(s): WGA, Wheat germ agglutinin. Why not use AP or HRP conjugated WGA. sections were prepared from decalcified onemonth - old mice femur and were used for IHC stain. In addition, the area of myocardial cells was quantified by staining with wheat germ agglutinin (WGA). For WGA/indirect immunouorescence double labelling, tissue sections were immunostained using either rabbit anti-desmoplakin I and II antiserum This staining was performed using standard protocols. In P. primaurelia soluble fluorescent wheat germ agglutinin, a lectin, is first accumulated below the plasmamembrane, followed by delivery to food vacuoles (Ramoino et al., 2001).Applying exogenous proxidase and EM analysis in P. multimicronucleatum reveals the We used a panel of seven dietary lectins to analyze cytotoxicity against hematological cancers. Compare Product No. According to the manual from Thermo, the WGA should be diluted in HBSS for staining. Thank you, Michael! I will now try to dilute the WGA with HBSS instead of PBS. Kind regards, Edward I use WGA Alexa Fluor 488 Conjugate on PFA fixxed and paraffin-embedded sections, and it works pretty well. It is diluted in PBS for a final concen The quantification of fibrotic tissue is an important task in the analysis of cardiac remodeling. Material Amount Storage* Stability Wheat germ agglutinin (WGA), fluorescein conjugate When CSF glycoproteins were stained with wheat germ agglutinin (WGA), the staining intensity was found to be significantly lower in the Alzheimer's disease group. Wheat germ agglutinin (WGA) (MW 36 kDa) is a dimeric, carbohydrate-free protein composed of two identical subunits. Description SDS Pricing; 61768: suitable for affinity chromatography: Afterwards, you can stain with additional antibodies. We therefore used Succinylated WGA is Hide. Hi Edward, what kind of WGA did you use? biotinylated or fluorophore labelled? Sometimes it helps to increase the concentration of the WGA or to pl Table 1. Paraffin sections of oral squamous cell carcinoma were stained with fluorescently labeled wheat germ agglutinin and with antibodies against structural proteins, which were revealed by immunohistochemistry with tyramide signal amplification. 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