Store at +4°C short term (1-2 weeks). NOT FOR USE IN DIAGNOSTIC PROCEDURES" For licensing inquiries, please contact partnerships@abcam.com, Rabbit polyclonal to Wheat Germ Agglutinin. Block nonspecific fluorophore binding sites with Blocking Buffer. After permeabilization, wash the tissue sections with three changes of PBSA-Triton Wash Buffer.

We recommended a WGA conjuage concentration of 5.0 µg/mL. Immediately mount the stained tissue with the selected mounting medium (do not allow the tissue to dry). … Cover the humidity chamber with aluminum foil to protect the fluorophores from light. Figure 8 . (C) Histological examination of cardiac hypertrophy by TRITC-WGA staining (upper). The SR staining marks the fibrotic tissue within the scar with a dark red dye; the remote myocardium, i.e. Agonists, activators, antagonists and inhibitors, Goat Anti-Rabbit IgG H&L (HRP) (ab205718), Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (, Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (, Suitable for: Immunodiffusion, Immunoelectrophoresis.

Publishing research using ab178444? Please contact us to place your order, or try again later. This protocol details a generalized procedure for staining tissue cryosections ranging from 5 to 20 micrometers in thickness. Goat Anti-Rabbit IgG H&L (HRP) (ab205718) When labeling is complete, remove the labeling solution, and wash the cells twice Slowly rotate the tissue sections as they are being permeabilized on an orbital shaker at 5-10 revolutions per minute. Immunocytochemistry applies to the study of the intracellular activities of proteins. Pixels with higher light intensities (brightness level 70 to 255) were only detected in the complete heart histogram. (B) the same section shown in A was co-stained with WGA-fluorescein (green) to outline general tissue structure. Dissolve the membranes in the tissue sections with Permeabilization Buffer by treatment for one or two hours for thin (5-10 micrometer) and thick (10-20 micrometer) sections, respectively.

Brightest point projection method to demonstrate the 3-dimensional appearance of capillaries in the heart, from the same z-stack as Fig. Return the stained slides to the staining jar and wash three times with PBSA-Triton Wash Buffer. I am trying to stain mice whole heart sections with WGA for cardiac myocyte size estimation and IB4 for microvascular density. 2.5 Wash cells. Custom antibody development and commercial partnerships to advance your diagnostic and therapeutic discovery. If incorrect, please enter your country/region into the box below, to view site information related to your country/region.

Place the slides in a humidity chamber (see Figure 2) and add the staining cocktail (250 to 500 microliters per slide) to the surface of the tissue section. The histogram analysis of WGA-FITC stained heart A (Figure 5C) shows a large overlap between background histogram (black border) and complete heart histogram (red border) in the range between brightness level 25 and 70. Guaranteed product quality, expert customer support. Please try the standard protocols listed below and let us know how you get on. Our Cookie Policy explains how you can opt-out of the cookies we use. Slowly rotate the tissue sections as they are being blocked on an orbital shaker at 5-10 revolutions per minute. Skip to Main Navigation. Cell membrane staining. Slowly rotate the tissue sections as they are being washed on an orbital shaker at 5-10 revolutions per minute. There are currently no Customer reviews or Questions for ab178444.Please use the links above to contact us or submit feedback about this product. Incubate the covered slides in the humidity chamber for one or two hours (thin and thick specimens, respectively) at 37 degrees. View complete list of fluorescent dye-conjugated lectins › Wheat germ agglutinin (WGA) is one of the most widely used lectins in cell biology. Primary - Upon delivery aliquot. WGA is found …

On the left (Panel A) DAPI appears in yellow/red, WGA-AF488 appears in green. WGA is a N acetylglucosamine specific lectin and is synthesized in developing and germinating embryos, roots and meristematic part of the leaves. Slowly rotate the tissue sections as they are being fixed on an orbital shaker at 5-10 revolutions per minute. 3 Heart Institute, Hadassah Hebrew University Medical Center, Jerusalem, Israel. If you continue without changing your cookie settings, we'll assume you’re happy with this. WGA is a protein found abundantly in wheat kernels. Protect from light with aluminum foil. Store at -20°C long term. On the right (Panel B) DAPI appears in blue, WGA-AF488 appears in green. Immunodiffusion, Immunoelectrophoresis, Secondary - the optimal conditions for tissue processing, staining, and microscope observations.
Hoechst (33342 and 33258) - 5 microliters in 150 milliliters of Hanks BSS (30 minutes). Counterstaining for nuclei using a variety of popular DNA-binding dyes follows treatment with the lectin and actin probes. The following resources may help you locate the website you're looking for: A | B | C | D | E | F | G | H | I | J | K | L | M | N | O | P | Q | R | S | T | U | V | W | X | Y | Z, 1000 E. University Ave. Laramie, WY 82071UW Operators (307) 766-1121 | Contact Us | Download Adobe Reader, Accreditation | Emergency Preparedness | Employment at UW | Gainfull Employment | Privacy Policy | Accessibility. Wash the counterstained specimens with either PBSA or Hanks Balanced Salt Solution (depending upon the nuclear dye) for three times at 5 minutes for each wash. 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. Immunohistochemistry (IHC) involves the study of cells and tissue.

This method of staining and analysis of cardiomyocytes enables accurate morphometric measurements in thick histological sections, thus unlocking the potential of archived tissue. After the final washing step, remove the slides from the staining jar and wipe excess buffer from the rear surface and edges of the slide. We use cookies to make our site as useful as possible. If you entered the URL manually, please check your spelling and try again. FWGA can easily … in the following tested applications.

4 answers. Scale bar, 0.5 cm. All rights reserved. Triticum vulgare Wheat germ agglutinin. DAPI - 5 microliters in 150 milliliters of PBSA (5 minutes). Scale bars, 100 or 50 μm as indicated. Among the useful fluorescent markers for visualization of tissues are rhodamine, fluorescein, the Alexa Fluor series, and the cyanine dyes. Fix 5-10 micrometer sections for 15-20 minutes and 10-20 micrometer sections for 20-30 minutes. The ratios of HW/BW and HW/TL (lower) (n = 6 per group). Each navigation link will open a list of sub navigation links. Nuclear Dye Wash Buffer - Hoechst and … ab178444 has not yet been referenced specifically in any publications. wheat germ agglutinin (WGA) selectively labels dermal extracellular matrix fibres and epidermal cell membranes in tissue sections of horse hoof lamellae, is compatible with indirect immunofluorescence and augments interpretation of indirect immunofluorescence antigen localization. Rabbit Anti-Wheat Germ Agglutinin antibody (ab178444) © 1998-2020 Abcam plc. When using Hoechst or SYTOX stains (30 minute incubation), first wash the slides in Hanks Balanced Salt Solution for two buffer exchanges prior to counterstaining. 2.4 Label cells. (C) combined image of CD31-Cy3 (red) and WGA-fluorescein (green) staining. Your browser does not have JavaScript enabled and some parts of this website will not work without it. WGA is a N acetylglucosamine specific lectin and is synthesized in developing and germinating embryos, roots and meristematic part of the leaves. Discussion. Carefully insert the frozen and mounted cryosections (on 1 x 3-inch microscope slides) into a vertical or horizontal staining jar. For thin sections (5-10 micrometers) block for one hour, and for thick sections (10-20 micrometers) block for two hours. Nuclear Dye Wash Buffer - Hoechst and SYTOX dyes require Hanks Balanced Salt Solution (Hanks BSS), while DAPI, as well as the monomeric and dimeric cyanine nuclear stains, can be used with PBSA. Align all sections to face in the same direction and avoid scratching the surface of the frozen tissue while loading the slides. Please note: All products are "FOR RESEARCH USE ONLY.

Lectin interaction with certain carbohydrates are as specific as enzyme to substrate or antigen to antibody interactions and have been used to study protein glycosylation. Please let us know so that we can cite the reference in this datasheet.

Permeabilization Buffer - 0.2 percent Triton X-100 in PBSA (sonicate detergent buffer for 30 minutes to one hour immediately before use). Apply a sufficient amount of labeling solution to cover cells adhering to coverslip(s). For thin sections (5-10 micrometers) wash for 5 minutes, and for thick sections (10-20 micrometers) wash for 10 minutes. The heart sections after SR (left image) and WGA-FITC staining (right image) are shown in Figure 2. Fluorescent WGA lectin co-staining of bone, ligament and tendon sections (A) a section of a palm from 3-day old rat was stained with antibodies against CD31-Cy3 (red) to visualize blood vessels. The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user. Question. To our knowledge, customised protocols are not required for this product.

The section was stained as detailed below using wheat germ agglutinin conjugated to Alexa Fluor 568 and phalloidin conjugated to Oregon Green 488. ab178444

Lectin interaction with certain carbohydrates are as specific as enzyme to substrate or antigen to antibody interactions and have been used to study protein glycosylation. (B) Representative images of H&E staining of the heart sections (upper). Sorry, we can't display this right now. IHC-P, WB, ELISA, IP. PBSA Wash Buffer - Use PBSA before permeabilization (after fixation) and after treatment with the phalloidin/lectin probes except for those tissues requiring specialized buffers for nuclear dyes that are not compatible with PBSA. It has a high binding affinity for N-acetyl-glucosamine (GlcNAc), one of the essential components of PG glycan strands, and thus can be used to probe PG specifically. Avoid freeze / thaw cycle. For DAPI and cyanine nuclear counterstains, add the diluted dye in PBSA to the staining jar and treat the specimen for the recommended time: 5-10 minutes for DAPI; 15-30 minutes for cyanine dyes (protect from light with aluminum foil). Presented in Figure 1 is a confocal image revealing striated actin fiber bundle structure in a thick (16 micrometer) section of rat tongue. WGA has been reported to have a role in dormancy and plant defence, but its characteristic biochemical properties and its predominance in growing and meristematic tissues, and its osmotic- and drought-stress inducibility suggest that this protein might be involved in a much wider range of functions. Nathan S. Claxton, Gregory K. Ottenberg, John D. Griffin, and Michael W. Davidson - National High Magnetic Field Laboratory, 1800 East Paul Dirac Dr., The Florida State University, Tallahassee, Florida, 32310.
Our Alexa Fluor® 488 conjugate of WGA exhibits the bright, green fluorescence of the Alexa Fluor® 488 dye (excitation/emission maxima ~495/519 nm). Images were collected in grayscale and subsequently pseudocolored with hues approximating the fluorescence emission spectra of the respective probes. For thin sections (5-10 micrometers) wash for 5 minutes, and for thick sections (10-20 micrometers) wash for 10 minutes.

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