Neurotensin Antibody

Neurotensin Antibody
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Price: $250.00
Product ID : 20072
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The ImmunoStar Neurotensin antiserum was quality control tested using standard immunohistochemical methods. The antiserum demonstrates strongly positive labeling of rat amygdala using indirect immunofluorescent and biotin/avidin-HRP techniques. Recommended primary dilution is 1/4000-1/8000 in PBS/0.3% Triton x-100 - Bn/Av-HRP Technique. Staining is completely eliminated by pretreatment with 10 µg of Neurotensin per 1 mL of diluted antibody.

Photo Description: IHC image of rat amygdala staining for neurotensin. The tissue was fixed with 4% formaldehyde in 0.1 M phosphate buffer, before being removed and prepared for vibratome sectioning. Floating sections were incubated at RT in 10% goat serum in PBS, before standard IHC procedure. Primary antibody was incubated at 1:5000 for 48 hours, goat anti-rabbit secondary was subsequently added for 1 hour after washing with PBS. Light microscopy staining was achieved with standard biotin-streptavidin/HRP procedure and DAB chromogen. 

Host: Rabbit

Quantity / Volume:  100 µL

State: Lyophilized Whole Serum

Reacts With: Cat, Dog, Frog, Human, Monkey, Mouse, Pigeon, Rat, Turtle

Availability: In Stock

Alternate Names:  neuromedin N; Preproneurotensin; pro-neurotensin

Gene Symbol / ID, Accession #:  NTS, 4922


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Journal References:

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Root, David H., et al. "Differential roles of ventral pallidum subregions during cocaine self‐administration behaviors." Journal of Comparative Neurology 521.3 (2013): 558-588. Tested in Rat, PubMed ID: 22806483

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Mumphrey, M. B., et al. "Roux‐en‐Y gastric bypass surgery increases number but not density of CCK‐, GLP‐1‐, 5‐HT‐, and neurotensin‐expressing enteroendocrine cells in rats." Neurogastroenterology & Motility 25.1 (2013): e70-e79. Tested in Rat, PubMed ID: 23095091

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Woulfe, J., et al. "Post-translational processing of the neurotensin/neuromedin N precursor in the central nervous system of the rat—II. Immunohistochemical localization of maturation products." Neuroscience 60.1 (1994): 167-181. Tested in Rat, PubMed ID: 8052410

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Berk, Mitchell L., Stacy E. Smith, and Harvey J. Karten. "Nucleus of the solitary tract and dorsal motor nucleus of the vagus nerve of the pigeon: Localization of peptide and 5‐hydroxytryptamine immunoreactive fibers." Journal of Comparative Neurology 338.4 (1993): 521-548. Tested in Pigeon, PubMed ID: 8132859

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Product Reviews

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Reviewed by droot
07/10/2012 - 10:12:41 PM
Visualization of ventromedial ventral pallidum
For our manuscript, Differential roles of ventral pallidum subregions during cocaine self-administration behaviors, in press at Journal of Comparative Neurology, we used the Immunostar rabbit anti-neurotensin primary antibody to visualize the ventromedial ventral pallidum subregion.

Rats were perfused with saline followed by 4% PF, stored in 30% sucrose, and coronally sectioned to 40 um. We followed the protocols of Zahm/Heimer and colleagues when they discovered the ventral pallidum subregions and their afferent/efferent projection patterns.

The protocol consisted of the following steps:
1. Wash in 0.1M phosphate buffer (PB; pH 7.4)
2. 15 min in 1% sodium borohydride
3. Wash
4. 1 hour blocking with PB containing 0.1% Triton X-100 and 3% normal goat serum
5. Primary antibody overnight at 4C - ImmunoStar rabbit anti-neurotensin diluted 1 : 6500 in PB containing 0.1% Triton X-100 and 3% normal goal serum
6. Wash
7. 1 hour anti-rabbit biotinylated secondary (Vector) 1:200 in PB with 0.1% Triton X-100
8. Wash
9. 1 hour ABC (Vector) 1:200 in PB with 0.1% Triton X-100
10. Wash
11. Develop with 0.05% DAB for 6 min

Our study involved recording neurons within the ventral pallidum subregions during specific aspects of intravenous cocaine self-administration (approaching toward, responding on, or retreating away from a cocaine-reinforced operandum). In order to verify the placement of microwires within the VP subregions, prior to perfusion we passed current through each stainless steel microwire to leave an iron deposit at the uninsulated tip. After DAB (brown reaction) and mounting, we visualized the iron deposit by incubating in a 5% potassium ferrocyanide and 10% HCl solution (leaving a blue-green reaction).

For our purposes, this antibody labeled fibers in ventromedial VP with low background. Individual neurons and other fibers were clearly observed within BNST that were not involved in our study.

I rated this antibody 4 stars because there was some variability for this stain between animals.

David H. Root, Ph.D.
Rutgers University
Reviewed by Dan_Tylee
04/05/2012 - 01:09:25 PM
Crisp, Cell-specific labeling
Tissue: Maccaca fasciscularis, perfused with saline and 4% paraformaldehyde, dehydrated through increase sucrose gradients, sectioned at 40 um with sliding microtome.

Method: Immunocytochemistry on free floating sections. Incubated with ImmunoStar's primary antibody at a concentration of 1:1000 for 4 days in 10% normal goat serum (in 0.1M PO4 buffer with .3% Triton X-100). Rinsed, then incubated with Vector Lab's biotinylated goat anti-rabbit secondary antibody (#BA-1000) at a concentration of 1:200 for 40 minutes at room temperature in the same 10% normal goat serum solution. Rinsed, then incubated with Vector's standard peroxidase kit (#PK-4000) for 60 minutes at room temperature in 0.1M PO4 buffer with .3% Triton X-100. Rinsed, then developed using the instructions for the kit.

Results: Crisp, cell-specific labeling. Low background. Very helpful for determining the boundary between the medial and lateral core divisions of the central nucleus of the amygdala.

Daniel Tylee
Department of Neurobiology & Anatomy
University of Rochester - School of Medicine and Dentistry