Alexa647 donkey anti rat

Manufactured by Jackson ImmunoResearch

Alexa647 donkey anti-rat is a secondary antibody conjugated with the fluorescent dye Alexa Fluor 647. This antibody is produced in donkeys and specifically binds to rat primary antibodies, allowing for their detection in various immunoassays and imaging applications.

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Lab products found in correlation

Mouse anti n cadherin 3b9, by Thermo Fisher Scientific (1 mentions) Rat anti pdgfrβ cd140b, by Thermo Fisher Scientific (1 mentions) Donkey anti rat cy3, by Jackson ImmunoResearch (1 mentions) Donkey anti mouse alexa 647, by Jackson ImmunoResearch (1 mentions) Prolong gold, by Thermo Fisher Scientific (1 mentions) Rat anti ha, by Roche (1 mentions) Lsm 700, by Zeiss (1 mentions) Donkey anti mouse cy3, by Jackson ImmunoResearch (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions) Bovine serum albumin solution, by Merck Group (1 mentions)

3 protocols using alexa647 donkey anti rat

Immunostaining of Muscle Cell Markers

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Immunostaining was performed with modifications from standard protocol previously described (Nielsen and Dymecki, 2010 (link)). Primary antibody dilutions: rat anti-CD13 (MCA2183GA) (1:500) (AbD Serotec); mouse anti-desmin (D33) (1:50) (DAKO/Agilent); mouse anti-N-cadherin (3B9) (1:300) (Invitrogen); rat anti-Pdgfrβ (CD140b) (1:50) (Invitrogen). Secondary antibodies: Cy3 donkey anti-rat; Alexa647 donkey anti-mouse; Alexa647 donkey anti-rat (all dilutions 1:500) (Jackson ImmunoResearch). For EdU incorporation assay, mice were injected intraperitoneally with 10 μg/gram of body weight with EdU at P5–7 or P8–10 or P12–14. On the day of harvest (P7 or P10 or P14), tissue was harvested 2 h post-injection. Detection of EdU used Click-iT™ Plus chemistry, with AlexaFluor^® 647 component following manufacturer’s instructions (Invitrogen). Cell nuclei were counterstained with 4′,6′-diamidino-2-phenylindole (DAPI). Tissue sections were mounted with ProLong Gold™ (Invitrogen) and coverslipped for imaging.

Selhorst S., Nakisli S., Kandalai S., Adhicary S, & Nielsen C.M. (2022). Pathological pericyte expansion and impaired endothelial cell-pericyte communication in endothelial Rbpj deficient brain arteriovenous malformation. Frontiers in Human Neuroscience, 16, 974033.

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Hippocampal Neuron Immunostaining Protocol

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Hippocampal neurons were transfected after 9 DIV as specified previously. At 12 DIV, neurons were incubated with rat anti-HA (Roche 3F10) for 30 min, then transferred into media containing Alexa 647 Donkey anti-Rat (Jackson Labs). After 30 min incubation in secondary antibodies, neurons were transferred back into conditioned media supplemented with fresh Neurobasal for 16 hours before being fixed, washed, and stained for proteins of interest. Co-aggregation studies were performed using multiple biological and technical replicates.

Steffen D.M., Ferri S.L., Marcucci C.G., Blocklinger K.L., Molumby M.J., Abel T, & Weiner J.A. (2021). The γ-Protocadherins interact physically and functionally with Neuroligin-2 to negatively regulate inhibitory synapse density and are required for normal social interaction. Molecular neurobiology, 58(6), 2574-2589.

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Vascularized Construct Imaging Protocol

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Constructs were fixed with paraformaldehyde (4%), for 20 min, and then permeabilized with 0.3% Triton X-100 (Bio-Lab Ltd.), for 10 min. Constructs were then washed with PBS and immersed overnight in Bovine serum albumin solution (5%; Millipore). Samples were then incubated with the following primary antibodies overnight at 4 °C: mouse anti–human-LYVE (1:200; R&D Systems), rat anti–mouse-LYVE (1:100; Santa Cruz), and anti-human desmin (1:50; Dako). Constructs were then treated with Alexa 647-donkey anti-rat (1:400) or Cy3-donkey anti-mouse (1:100; Jackson Immunoresearch Laboratory) secondary antibodies and DAPI (Sigma-Aldrich) for 3 h, at room temperature. Evans blue was injected into the vascularized construct using an insulin needle, followed by 10 min incubation, after which constructs were imaged.
Whole vascularized constructs were imaged with a confocal microscope (LSM700, Zeiss), using 2.5×, 5×, 20×, and 63× oil immersion lenses. All image analyses were quantified using self-written algorithms in MATLAB that transformed captured images into binary images prior to analysis.

Landau S., Newman A., Edri S., Michael I., Ben-Shaul S., Shandalov Y., Ben-Arye T., Kaur P., Zheng M.H, & Levenberg S. (2021). Investigating lymphangiogenesis in vitro and in vivo using engineered human lymphatic vessel networks. Proceedings of the National Academy of Sciences of the United States of America, 118(31), e2101931118.

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