At 36 hours after transfection, Cos7 cells expressing FLAG-NgR1 and HA-ORL1 were fixed with 4% paraformaldehyde for 15 min and then permeabilized with 0.1% Triton X-100 in phosphate-buffered saline (PBS) for 15 min. The samples were incubated overnight at 4°C with anti-FLAG (1:1000) and anti-HA (1:1000), or anti-NgR1 (1:1000), anti-HA (1:1000), and anti-GM130 (1:500) antibodies. Then, either Alexa 488–conjugated donkey anti-mouse immunoglobulin G (IgG) and Alexa 568–conjugated donkey anti-rabbit IgG, or Alexa 488–conjugated donkey anti-goat IgG, Alexa 568–conjugated donkey anti-rabbit IgG, and Alexa 647–conjugated donkey anti-mouse IgG (1:2000; all from Invitrogen) were used to detect primary antibodies. Samples were mounted with mounting solution (Vector Laboratories) and observed using an LSM710 confocal microscope.
Alexa 568 conjugated donkey anti rabbit igg
Manufactured by Thermo Fisher Scientific
Sourced in United States
Alexa Fluor 568–conjugated donkey anti-rabbit IgG is a secondary antibody reagent used for detecting and visualizing rabbit primary antibodies in various immunodetection techniques, such as Western blotting, immunohistochemistry, and flow cytometry. The Alexa Fluor 568 fluorescent dye is conjugated to the donkey anti-rabbit IgG, allowing for sensitive and specific detection of target proteins or cellular structures.
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Lab products found in correlation
Prolong gold antifade reagent, by Thermo Fisher Scientific (1 mentions)
Mounting solution, by Vector Laboratories (1 mentions)
Alexa 488 conjugated donkey anti goat igg, by Thermo Fisher Scientific (1 mentions)
Lsm710 confocal microscope, by Vector Laboratories (1 mentions)
Alexa 647 conjugated donkey anti mouse igg, by Thermo Fisher Scientific (1 mentions)
2 protocols using alexa 568 conjugated donkey anti rabbit igg
1
Visualizing NgR1 and ORL1 Receptor Localization
2
Immunohistochemical Visualization of Midbrain Neurons
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Glass slides containing serial VTA and NAc sections were first blocked for 3–4 h and then incubated overnight at 4°C with a mixture of the following antibodies: mouse anti-NeuN (1:500; IgM; Millipore Bioscience Research Reagents, Merk, USA) and rabbit anti-TH (1:1,000; Millipore Bioscience Research Reagents, Merk, USA). After incubation with primary antibodies and subsequent washing with PB, sections were incubated in a mixture of secondary antibodies: Alexa 488 conjugated donkey anti-mouse IgM and Alexa 568-conjugated donkey anti-rabbit IgG (1:500; Invitrogen, ThermoFisher Scientific, Waltham, MA, USA) for 2 h. Sections were mounted in superfrost plus gold slides (ThermoFisher Scientific, Waltham, MA, USA), air-dried, and coverslipped with ProLong™ Gold antifade reagent (Invitrogen, ThermoFisher Scientific, Waltham, MA, USA).
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