Rabbit anti neuropeptide y

Manufactured by Immunostar

Rabbit anti-neuropeptide Y is a primary antibody that recognizes neuropeptide Y, a regulatory peptide found in the central and peripheral nervous systems. It is commonly used in research applications to detect and quantify neuropeptide Y expression in biological samples.

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2 protocols using rabbit anti neuropeptide y

Immunofluorescent Staining of Neuronal Markers

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Free-floating fixed sections were blocked in 5% goat serum, 0.5% Triton X-100 (TX) in PBS for 1 hour at room temperature, and then incubated with primary antibody in 1.25% goat serum, 0.125% TX in PBS for 48 hours at 4°C. Sections were then washed 3x with PBS and incubated with secondary antibodies in 1% goat serum, 0.1% TX in PBS for 2 hours at room temperature. Finally, sections were washed 3x with PBS and mounted in Vectashield Antifade (Vector Laboratories) or Prolong Diamond (Molecular Probes) mounting medium. Sections stained with mouse primary antibodies were blocked in ReadyProbes “Mouse on Mouse” IgG blocking solution (Thermofisher) for 1 hour before the initial blocking step. Primary antibodies and dilutions used were rabbit anti-calretinin (Swant, 1:2000), mouse anti-calbindin (Swant, 1:500), and rabbit anti-neuropeptide Y (Immunostar, 1:2000). Secondary antibodies used were AlexaFluor 647 anti-rabbit (Thermofisher, 1:1000) and AlexaFluor 647 anti-mouse (Thermofisher, 1:500).

Hostetler R.E., Hu H, & Agmon A. (2023). Genetically Defined Subtypes of Somatostatin-Containing Cortical Interneurons. bioRxiv.

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Immunohistochemical Analysis of Brain Tissue

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Tissue preparation and immunohistochemistry were done as described (Palop et al., 2011 (link)). After fixation, brains were cryoprotected in 30% sucrose in phosphate-buffered saline (PBS) for 2 days and sectioned with a sliding microtome (Leica Microsystems) into 30-µm sections. Ten stereological subseries sections from each hemibrain were stored in a solution containing 30% glycerol, 30% ethylene glycol, and 40% PBS at −20°C until staining. Primary antibodies used included rabbit anti-GFP (1:3,000; Life Technologies), chicken anti-GFP (1:500; Aves Labs), rabbit anti-Nav1.1 (1:500; Alomone), mouse anti-parvalbumin (1:3,000; Swant), rabbit anti-parvalbumin (1:3,000; Swant), rat anti-somatostatin (1:50; Novus), rabbit anti-neuropeptide Y (1:2000; Immunostar), rabbit anti-Iba-1 (1:1000; Wako), rabbit anti-GFAP (1:3000; Sigma), and biotinylated 82E1 (1:1000; Immuno-Biological Laboratories). Primary antibodies were detected with biotinylated donkey anti-rabbit (1:500; Jackson ImmunoResearch), Alexa Fluor 488 donkey anti-rabbit (1:300; Life Technologies), Alexa Fluor 488 donkey anti-chicken (1:300; Life Technologies), Alexa Fluor 594 donkey anti-rat (1:300; Life Technologies), Alexa Fluor 594 donkey anti-mouse (1:300; Life Technologies), or Alexa Fluor 594 donkey anti-rabbit (1:300; Life Technologies).

Martinez-Losa M., Tracy T.E., Ma K., Verret L., Clemente-Perez A., Khan A.S., Cobos I., Ho K., Gan L., Mucke L., Alvarez-Dolado M, & Palop J.J. (2018). Nav1.1-Overexpressing Interneuron Transplants Restore Brain Rhythms and Cognition in a Mouse Model of Alzheimer’s Disease. Neuron, 98(1), 75-89.e5.

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