Alexa 488 conjugated donkey anti rabbit secondary antibody

Manufactured by Thermo Fisher Scientific

Sourced in United States

The Alexa Fluor 488-conjugated donkey anti-rabbit secondary antibody is a detection reagent used in immunoassays and immunofluorescence experiments. It is designed to bind to primary rabbit antibodies, allowing for the visualization of target proteins or other biomolecules.

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Lab products found in correlation

Lm710 confocal microscope, by Zeiss (1 mentions) M o m kit, by Vector Laboratories (1 mentions) Vectamount, by Vector Laboratories (1 mentions) Anti lyve 1 antibody, by Abcam (1 mentions) Hanks balanced salt solution (hbss), by Merck Group (1 mentions) Annexin 5 apc, by BD (1 mentions)

Close spelling variants of this product

Alexa Fluor 488-conjugated donkey anti-rabbit secondary antibody Alexa Fluor 488-conjugated donkey anti rabbit IgG secondary antibody Alexa 488-conjugated donkey anti-rabbit IgG secondary antibody Donkey anti-rabbit Alexa 488 secondary antibody Alexa-488 conjugated donkey anti-rabbit antibody Alexa-488 conjugated anti-rabbit secondary antibody Alexa rabbit 488 conjugated secondary antibody Alexa 488-conjugated donkey anti-rabbit Anti-rabbit Alexa-488 secondary antibody Alexa 488-conjugated anti-rabbit antibody

4 protocols using alexa 488 conjugated donkey anti rabbit secondary antibody

Immunofluorescence Imaging of DDX1 Protein

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Cells cultured on coverslips were fixed and processed as previously described [15 (link),35 ]. Cells were incubated with a rabbit anti-DDX1 antibody (batch 2923; 1:1000 dilution) [35 ,39 (link)] followed by Alexa 488-conjugated donkey anti-rabbit secondary antibody (Life Technologies). Coverslips were mounted onto slides in polyvinyl alcohol (Calbiochem)-based mounting medium containing 1 μg/ml 4′,6′-diamidino-2-phenylindole (DAPI). Images were acquired using a Zeiss LM710 confocal microscope, exported as TIFF files using ZEN and assembled using Photoshop software.

Garg M., Li L, & Godbout R. (2023). Role of DDX1 in the oxidative response of ataxia telangiectasia patient-derived fibroblasts. Redox Biology, 69, 102988.

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Muscle Morphology and Fibrosis Analysis

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Tissue sections (10 μm thick) were cut and stained with hematoxylin and eosin for the assessment of overall muscle morphology. Collagen fibers were detected by sirius red staining. Sections were fixed by incubation for 10 min with 4% formaldehyde and dried. They were then incubated with a 0.3% solution of sirius red in aqueous saturated picric acid for 1 h, washed in acidified water (0.5% acetic acid), dehydrated, and mounted in VectaMount (Vector Laboratories, Peterborough, UK). Fibrosis was then quantified in a nonblind manner by determining the proportion of the total cross-sectional area stained red with ImageJ software.
We then assessed dystrophin levels on cryosections by incubation with the Lab Vision Dystrophin (1:500; Thermo Scientific, Waltham, MA, USA) rabbit polyclonal antibody and β-sarcoglycan expression by incubation with a monoclonal antibody against this protein (1:50; Novocastra, Newcastle, UK). The binding of primary antibodies was detected by incubation with an Alexa 488-conjugated donkey anti-rabbit secondary antibody (1:1,000; Life Technologies, Carlsbad, CA, USA) and an Alexa 594-conjugated goat anti-mouse secondary antibody (1:1,000; Life Technologies, Carlsbad, CA, USA). The monoclonal antibody was used with the M.O.M. kit (Vector Laboratories, Peterborough, UK).

Forand A., Muchir A., Mougenot N., Sevoz-Couche C., Peccate C., Lemaitre M., Izabelle C., Wood M., Lorain S, & Piétri-Rouxel F. (2020). Combined Treatment with Peptide-Conjugated Phosphorodiamidate Morpholino Oligomer-PPMO and AAV-U7 Rescues the Severe DMD Phenotype in Mice. Molecular Therapy. Methods & Clinical Development, 17, 695-708.

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Visualizing Mitochondrial Morphology in U2OS Cells

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U2OS cells were cultured on glass bottomed dishes (MatTek, Cat. # P35G-1.5–14C) at 37 C and 5% CO₂. Prior to labeling, cells were rinsed 3 times with 1X PBS, fixed with 1 mL paraformaldehyde/glutaraldehyde (4%/2%) (Electron Microscopy Sciences, Cat. # 15710 and 16120) in 1X PBS for 20 minutes at 37C, rinsed twice in 2 mL 750 mM Tris-HCL pH 7.5 (Corning, Cat. # 46–030-CM), and permeabilized in 0.2% Triton-X (Sigma, Cat. # T9284)/1X PBS for 10 minutes. Next, samples were washed 3 times in staining buffer (0.2% Tween-20 (Sigma, Cat. # P9416)/1X PBS) and blocked in staining buffer containing 1% bovine serum albumin (BSA, Thermo Fisher Scientific, 37525) for 30 minutes. The blocking buffer was removed, and the samples stained with 200 μL of 1:200 anti-Tomm20 primary antibody (Abcam, Cat. # 78547): 0.2% Tween-20/1X PBS for 1 hour. Cells were washed in 0.2% Tween-20/1X PBS and stained with 200 μL of 1:200 Alexa-488 conjugated donkey anti-rabbit secondary antibody (Invitrogen, Cat. # A21206) for 1 hour. Finally, cells were washed 3 times in 0.2% Tween-20/1X PBS and imaged in the instant SIM in 1X PBS.

Guo M., Li Y., Su Y., Lambert T., Nogare D.D., Moyle M.W., Duncan L.H., Ikegami R., Santella A., Rey-Suarez I., Green D., Beiriger A., Chen J., Vishwasrao H., Ganesan S., Prince V., Waters J.C., Annunziata C.M., Hafner M., Mohler W.A., Chitnis A.B., Upadhyaya A., Usdin T.B., Bao Z., Colόn-Ramos D., La Riviere P., Liu H., Wu Y, & Shroff H. (2020). Rapid image deconvolution and multiview fusion for optical microscopy. Nature biotechnology, 38(11), 1337-1346.

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Isolation and Analysis of Lymphatic Endothelial Cells

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Cells isolated from E15.5 control (Notch1^flx/flx) and conditional Notch1 mutant (Notch1^flx/flx;Prox1CreER^T2) embryos were stained for Lyve-1 and CD31 and subjected to FACS. Briefly, E15.5 embryos were harvested in Hank’s balanced salt solution (HBSS, Sigma-Aldrich) and then chopped for an overnight digestion with collagenease I/II. The colleagenase-treated cell suspension was incubated with RBC (Red blood cell) lysis buffer (StemCell Technologies). Following centrifugation, cell pellets were incubated with anti-Lyve-1 antibody (Abcam) for 20 min at 4°C. After washing with PBS, the cells were then stained with PE-conjugated anti-CD31 antibody (BD Pharmingen) and Alexa 488-conjugated donkey anti-rabbit secondary antibody (Invitrogen). After gauze filtration with a cell strainer (40μm BD Biosciences) to obtain a single cell suspension, the cells were subjected to staining with Annexin V - APC (BD Pharmingen) to assess cell death in the sorted LECs using Flow Fortessa.

Fatima A., Culver A., Culver F., Liu T., Dietz W.H., Thomson B.R., Hadjantonakis A.K., Quaggin S.E, & Kume T. (2014). Murine Notch1 is required for lymphatic vascular morphogenesis during development. Developmental dynamics : an official publication of the American Association of Anatomists, 243(7), 957-964.

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