Wheat germ agglutinin alexa fluor 488

Manufactured by Thermo Fisher Scientific

Sourced in United States

Wheat Germ Agglutinin Alexa Fluor 488 is a fluorescently labeled lectin that binds specifically to N-acetylglucosamine and sialic acid residues on cell surfaces. It is used as a fluorescent probe for the detection and visualization of these carbohydrate moieties in biological samples.

Automatically generated - may contain errors

Lab products found in correlation

Spelling variants of this product

Alexa Fluor 488 (6806 citations) Alexa 488 (1863 citations) Alexa Fluors (21 citations) Wheat germ agglutinin‐Alexa Fluor 488 conjugate (5 citations) Wheat Germ Agglutinin-Alexa Fluor® 488 conjugate (WGA-Alexa Fluor® 488) (3 citations) Alexa Fluor 488 wheat germ agglutinin (WGA, (2 citations) Alexa Fluor 488® conjugate of wheat germ agglutinin (WGA) solution (2 citations) Wheat Germ Agglutinin (WGA) Alexa Fluor 488 and 350 dyes (2 citations) Wheat Germ Agglutinin Alexa Fluor 488 Conjugate (WGA, (2 citations) Alexa Fluor 488-conjugated wheat germ agglutinin (WGA; (2 citations)

20 protocols using wheat germ agglutinin alexa fluor 488

Fluorescent Tracer Uptake in Nephrocytes

Check if the same lab product or an alternative is used in the 5 most similar protocols

Fluorescent tracer uptake in nephrocytes to evaluate nephrocyte function was performed as previously described (Hermle et al., 2017 (link)). Briefly, nephrocytes were dissected in PBS and incubated with FITC-albumin (#A9771, Sigma) for 30 s. After a fixation step of 5 min in 8% paraformaldehyde, cells were rinsed in PBS and exposed to Hoechst 33342 (1:1000, #H1399, Thermofisher) for 20 s and mounted in Roti-Mount (#HP19.1, Carl Roth). Cells were imaged using a Zeiss LSM 880 laser scanning microscope. Quantitation of fluorescent tracer uptake was performed with ImageJ software. The results are expressed as a ratio to a control experiment with flies carrying the (heterozygous) GAL4 transgene but no UAS that was performed in parallel.
The parallel recording of two fluorescent tracers of different size to study the passage of the slit diaphragm was carried out in the same way as the assay for nephrocyte function, except that nephrocytes were simultaneously incubated with tracers FITC-albumin (0.2 mg/ml) and Texas-Red-Dextran (#D1863, Thermofisher, 10 kDa, 0.2 mg/ml) for 30 s after dissection. Cells were imaged using a Zeiss LSM 880 laser scanning microscope. Image quantitation was performed with ImageJ software for each channel separately. Alternative tracers were Texas-Red-Avidin (66 kDa, #A2348, Sigma) and Alexa Fluor 488 wheat germ agglutinin (38 kDa, #W11261, Thermofisher).

Lang K., Milosavljevic J., Heinkele H., Chen M., Gerstner L., Spitz D., Kayser S., Helmstädter M., Walz G., Köttgen M., Spracklen A., Poulton J, & Hermle T. (2022). Selective endocytosis controls slit diaphragm maintenance and dynamics in Drosophila nephrocytes. eLife, 11, e79037.

+ Open protocol

+ Expand

Nanoparticle Uptake in RAW 264.7 Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

RAW 264.7 cells (ATCC) were maintained in Dulbecco’s Modified Eagles Medium (DMEM, Invitrogen) supplemented with 10% fetal calf serum (Atlanta Biologicals), 100 IU penicillin (Invitrogen), 100 μg/mL streptomycin (Invitrogen), and 200 mM L-glutamine (Invitrogen). For examination of nanoparticle uptake, cells were seeded at a density of 10×10³ cells/well in optical-bottom 96-well plates (Ibidi, 89626) in phenol red free media. After 24 hours in culture, cells were incubated with CDNP-VT680 (50 μg/mL) for the specified time points, washed by PBS, and fixed with paraformaldehyde (4%, 30 min, 37°C). Cells were subsequently stained for cell membrane (5.0 μg/mL Alexa Fluor 488 wheat germ agglutinin, Thermo Fisher) and nuclei (DAPI, Invitrogen) for 15 min at room temperature. For lysosome imaging, cells were similarly stained via DAPI and Alexa Fluor 555 wheat germ agglutinin, permeabilized by 0.2% Triton X-100 and 2.25% BSA in PBS, and incubated with anti-LAMP1 Alexa Fluor 488. Plates were washed and subsequently imaged on a custom high-content screening microscope (Olympus). Acquired images were imported into CellProfiler (Broad Institute) for automated segmentation and analysis of nanoparticle uptake: quantified as integrated fluorescent intensity per cell, normalized to the mean intensity at 24 hours.

Ahmed M.S., Rodell C.B., Hulsmans M., Kohler R.H., Aguirre A.D., Nahrendorf M, & Weissleder R. (2019). A Supramolecular Nanocarrier for Delivery of Amiodarone Anti-Arrhythmic Therapy to the Heart. Bioconjugate chemistry, 30(3), 733-740.

+ Open protocol

+ Expand

Proximity Ligation Assay for Immune Cell Signaling

Check if the same lab product or an alternative is used in the 5 most similar protocols

Immunofluorescence analysis of RBL-2H3 cells and BMMC was performed as described (80) . PLA was performed using a Duolink in situ kit (Sigma-Aldrich). Cultured BMMCs (100,000 cells) were plated on coverslips coated with fibronectin (10 g/ml; Sigma-Aldrich) for 45 min in MnCl 2 (1 mM)-containing medium and maintained at 37°C in a humidified atmosphere. After PMA/ionomycin (20 nM:1 M) stimulation for indicated time points, cells were fixed, Downloaded from https://www.science.org on September 06, 2024 quenched, permeabilized (16) , and blocked for 1 hour at 37°C before incubation with primary Abs for 2 hours at room temperature. PLA was then performed according to the manufacturer's instructions. Complexes were analyzed using an LSM 780 Zeiss confocal microscope (Carl Zeiss) at 63× steps of a 0.38-m microscope. Images were processed with Imaris 8.1 software (Bitplane, Oxford Instruments), and the spots per nucleus were quantified. To evaluate membrane or cytoplasmic localization, cells were additionally labeled with Alexa Fluor 488-wheat germ agglutinin (Thermo Fisher Scientific) before the permeabilization procedure, and dots were evaluated for membrane-proximal or cytoplasmic localization using Imaris 8.1 software counting at least 30 cellular sections.

Madera-Salcedo I.K., Danelli L., Tiwari N., Dema B., Pacreau E., Vibhushan S., Birnbaum J., Agabriel C., Liabeuf V., Klingebiel C., Menasche G., Macias-Silva M., Benhamou M., Charles N., González-Espinosa C., Vitte J, & Blank U. (2018). Tomosyn functions as a PKCδ-regulated fusion clamp in mast cell degranulation. Science signaling, 11(537).

+ Open protocol

+ Expand

Quantifying Muscle Fiber Morphology

Check if the same lab product or an alternative is used in the 5 most similar protocols

10-μm TA sections were blocked in 5% BSA (Fisher Scientific), stained with Alexa Fluor 488 wheat germ agglutinin (WGA) (Molecular Probes, Invitrogen Detection Technologies), and mounted in Vectashield Hard Set with DAPI (Vector Laboratories, Burlingame, CA). Images were captured using a Zeiss Axioskop 2 Plus fluorescence microscope, a Zeiss AxioCam HRc digital camera, and Axiovision 4.8 software or an Olympus FluoviewFV1000 laser-scanning biological confocal microscope using the Olympus Micro FV10-ASW 3.1 software. CLN percentage and Feret’s minimal diameter were measured after stitching together images to recreate a montage of the entire TA muscle section.

Wuebbles R.D., Cruz V., Van Ry P., Barraza-Flores P., Brewer P.D., Jones P, & Burkin D.J. (2019). Human Galectin-1 Improves Sarcolemma Stability and Muscle Vascularization in the mdx Mouse Model of Duchenne Muscular Dystrophy. Molecular Therapy. Methods & Clinical Development, 13, 145-153.

+ Open protocol

+ Expand

Multicolor Immunofluorescence Microscopy

Check if the same lab product or an alternative is used in the 5 most similar protocols

Immunofluorescence and microscopy were undertaken as previously described [8 (link)]. Briefly, a TRITC directly conjugated hBCL-2 (Santa Cruz Biotechnology, Dallas, TX, USA), an anti-NRAS monoclonal antibody was visualized with a goat anti-mouse Alexa 647 secondary antibody and anti-mitochondria antibody Tom 20 (Santa Cruz Biotechnology, Dallas, TX, USA) visualized with a goat anti-rabbit Alexa 488 secondary antibody. The fluorescent lectin (Alexa Fluor 488) wheat germ agglutinin (Molecular Probes, Invitrogen, Paisley, OR, USA) was used as a plasma membrane marker for mouse cells. Slides were analyzed by confocal microscopy on a Zeiss LSM 510 META confocal laser microscope (Zeiss, Jena, Germany).

Gorombei P., Guidez F., Ganesan S., Chiquet M., Pellagatti A., Goursaud L., Tekin N., Beurlet S., Patel S., Guerenne L., Le Pogam C., Setterblad N., de la Grange P., LeBoeuf C., Janin A., Noguera M.E., Sarda-Mantel L., Merlet P., Boultwood J., Konopleva M., Andreeff M., West R., Pla M., Adès L., Fenaux P., Krief P., Chomienne C., Omidvar N, & Padua R.A. (2021). BCL-2 Inhibitor ABT-737 Effectively Targets Leukemia-Initiating Cells with Differential Regulation of Relevant Genes Leading to Extended Survival in a NRAS/BCL-2 Mouse Model of High Risk-Myelodysplastic Syndrome. International Journal of Molecular Sciences, 22(19), 10658.

+ Open protocol

+ Expand

Tracking Cell Division and Lineages

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

To track the in vivo growth of dividing single cells and lineages, bacteria were tracked by time-lapse microscopy. Cells were grown on agar pads (64 (link), 67 (link)) containing 15 mg ml⁻¹ of electrophoresis-grade agarose (Fisher Scientific) in M9 medium with the desired concentration of streptomycin. All cells used to inoculate agar pads were grown exponentially in streptomycin-free M9 medium. Time-lapse images were capture on a Nikon Eclipse Ti microscope with the Nikon NIS Element AR software by 100× phase-contrast and fluorescence (Prior, Lumen 200) imaging. To visualize the distribution of cell wall during the formation of minicells, cells were pulse-labeled with Alexa Fluor 488 wheat germ agglutinin (WGA) (Life Technologies Corporation) (68 (link)). Phase-contrast images were taken at intervals of 2 min. To minimize phototoxicity and bleaching, fluorescence images were recorded at longer intervals of 10 and 30 min. Microscope and agar pads were kept at 37°C with a Nevtek ASI 400 heater.

Rang C.U., Proenca A., Buetz C., Shi C, & Chao L. (2018). Minicells as a Damage Disposal Mechanism in Escherichia coli. mSphere, 3(5), e00428-18.

+ Open protocol

+ Expand

Detecting Protein Expression via RNAscope and Lectin Labeling

Check if the same lab product or an alternative is used in the 5 most similar protocols

In situ hybridization was conducted with RNAscope® 2.0 HD Assay (red) probes for mouse bfsp2 (CP49) prepared by Advanced Cell Diagnostics. Tissue was processed per the manufacturer’s instructions. To test whether lectin labeling was significantly affected, sections were labeled with Alexa Fluor 488-wheat germ agglutinin (WGA; Life Technologies, Cat# W11261).

Sun N., Shibata B., Hess J.F, & FitzGerald P.G. (2015). An alternative means of retaining ocular structure and improving immunoreactivity for light microscopy studies. Molecular Vision, 21, 428-442.

+ Open protocol

+ Expand

Measuring Cardiomyocyte Cross-Sectional Areas

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Whole rat hearts were excised from the euthanized animals and wet heart weights were measured on an OHAUS electronic balance machine (NJ, USA). Heart gravimetric indices were determined by calculating heart weight (mg)-to-body weight (g) ratios and heart weight (mg)-to-tibia length (cm) ratios. To assess cardiomyocytes cross-sectional areas, formalin fixed, paraffin embedded 5 μm thick rat heart LV tissue sections were deparaffinized, hydrated, heat-induced antigen retrieved and stained with Alexa Fluor 488 Wheat Germ Agglutinin (5 μg/mL, Invitrogen) for 1 h at room temperature (RT). 4',6-diamidino-2-phenylindole (DAPI, Invitrogen) was used to counterstain nuclei [23 (link),42 ,43 ]. We used Nikon A1R high-resolution confocal microscope (Nikon Instruments inc., Melville, NY) coupled with Nikon NIS Elements Software (v4.13.04) to image the stained tissue sections on a 60× oil objective lens (NA = 1.4). Cardiomyocytes cross sectional areas (μm²) were measured in an analyst-blinded manner on alphanumerically labeled images using NIH ImageJ software (Bethesda, MD) [23 (link)].

Abdullah C.S., Remex N.S., Aishwarya R., Nitu S., Kolluru G.K., Traylor J., Hartman B., King J., Bhuiyan M.A., Hall N., Murnane K.S., Goeders N.E., Kevil C.G., Orr A.W, & Bhuiyan M.S. (2022). Mitochondrial dysfunction and autophagy activation are associated with cardiomyopathy developed by extended methamphetamine self-administration in rats. Redox Biology, 58, 102523.

+ Open protocol

+ Expand

Cardiomyocyte Cross-Sectional Area Measurement

Check if the same lab product or an alternative is used in the 5 most similar protocols

Briefly, paraffin‐embedded heart tissue sections (5 μmol/L) were dewaxed and hydrated, and antigen was retrieved by boiling at 100°C in 10 mmol/L sodium citrate buffer (pH 6) for 30 minutes.52 The sections were then blocked with blocking solution (1% BSA, 0.1% cold water fish skin gelatin, and 1% Tween 20 in PBS) for 1 hour at room temperature followed by incubation with Alexa Fluor 488 wheat germ agglutinin (5 μg/mL, Invitrogen) and 4′,6‐diamidino‐2‐phenylindole (Invitrogen) for 1 hour at room temperature. The slides were then washed with PBS and mounted with Vectashield Hard Set (Vector Laboratories, Burlingame, CA). Stained tissue sections were observed and evaluated by an investigator blinded to genotype for wheat germ agglutinin staining on a Leica TCS SP5 spectral confocal microscope using a ×63 oil objective lens where images were acquired with Leica LAS (AF 2.6.3) software. All cardiomyocyte cross‐sectional areas were measured in each ×63 confocal field to calculate the average size of cardiomyocytes in ImageJ (v1.49) software (NIH).58

Abdullah C.S., Alam S., Aishwarya R., Miriyala S., Panchatcharam M., Bhuiyan M.A., Peretik J.M., Orr A.W., James J., Osinska H., Robbins J., Lorenz J.N, & Bhuiyan M.S. (2018). Cardiac Dysfunction in the Sigma 1 Receptor Knockout Mouse Associated With Impaired Mitochondrial Dynamics and Bioenergetics. Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease, 7(20), e009775.

+ Open protocol

+ Expand

Bacterial Culture and Microscopic Analysis

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Incubation of S. mutans, S. aureus, and S. pyogenes in liquid media: To ensure a pure culture, fresh S. mutans, S. aureus, and S. pyogenes from agar plates were inoculated in liquid media and cultured at 37 °C with 5% carbon dioxide in the incubator one day prior to an experiment.
Capturing, fixing, and staining bacteria: The procedures for capturing, fixing, and staining bacteria in liquid media are detailed in our previous paper.^{1 (link)} In brief, after culturing overnight, the bacteria suspensions were homogenized with vortexing and added to each CandyCollect device at a volume of 50 μL (devices negative controls were loaded with 50 μL of PBS). Bacteria were incubated in the device for 10 min. For devices that were imaged, bacteria were fixed with 4% paraformaldehyde (PFA) for 15 min, and 50 μL of Alexa Fluor^™ 488 Wheat Germ Agglutinin (WGA, Invitrogen^™, Fisher Scientific, Cat# W11261, 1 mg/mL) at 1:500 dilution (v/v) was added to the channel for staining S. aureus and S. pyogenes; 50 μL of 1:200 (v/v) WGA was added for staining S. mutans. An additional three devices were evaluated for a mixture of S. mutans, S. aureus and S. pyogenes, each at a concentration of 10⁴ CFU/mL to match physiological bacterial concentration for detection of bacteria in a mixture.

Tu W.C., McManamen A.M., Su X., Jeacopello I., Takezawa M.G., Hieber D.L., Hassan G.W., Lee U.N., Anana E.V., Locknane M.P., Stephenson M.W., Shinkawa V.A., Wald E.R., DeMuri G.P., Adams K., Berthier E., Thongpang S, & Theberge A.B. (2023). At-home saliva sampling in healthy adults using CandyCollect, a lollipop-inspired device. Analytical chemistry, 95(27), 10211-10220.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!