Substance P Antibody

Substance P Antibody
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Price: $275.00
Product ID : 20064

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Description

The ImmunoStar Substance P antiserum was quality control tested using standard immunohistochemical methods. The antiserum demonstrates strongly positive labeling of rat substantia nigra and spinal cord using biotin/avidin-HRP techniques. Recommended primary dilution is 1/6000-1/8000 in PBS/0.3% Triton X-100 - biotin/avidin-HRP Technique.

The specificity of the antiserum for Substance P was demonstrated using soluble pre-adsorption with the peptides in question at a final concentration of 10 µg of peptide per mL of diluted antiserum. Substance P immunolabeling was completely abolished by pre-adsorption with Substance P. Pre-adsorption with the following peptides resulted in no reduction of immunostaining: neurokinin A, neurokinin B, somatostatin and neuropeptide K.

Photo Description: IHC image of rat spinal cord (above) and rat tegmentum (below) staining for substance p. The tissue was fixed with 4% formaldehyde in phosphate buffer, before being removed and prepared for vibratome sectioning for brain and frozen sectioning for spinal cord. 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.

Substand P tegmentum #20064

Host: Rabbit

Quantity / Volume:  100 µL

State: Lyophilized Whole Serum

Reacts With: Bird, Bullfrog, Cat, Chick, Chicken, Dog, Dolphin, Doves, Equine, Fish, Frog, Gerbil, Goat, Goldfish, Guinea Pig, Hamster, Horse, Human, Monkey, Mouse, Mudpuppy, Narwhal, Parrot, Pig, Pig, Pigeon, Quail, Rabbit, Ram (Sheep), Rana Pipiens (Leopard Frog), Rat, Ratfish, Salmon, Scorpaena Porcus (Fish), Sea Lamprey, Sheep, Snail, Sparus Aurata (Fish), Squalus Acanthias (Dogfish), Squirrel, Starfish, Sting Ray, Sturgeon (Fish), Three-Spined Stickleback (Fish), Trout, Turtle, Vole, Zebra Finch, Zebrafish

Availability: In Stock

Alternate Names:  Protachykinin-1; PPT; tachykinin peptide hormone; NKA, NKNA, TAC2; tachykinin, precursor 1, anti-substance P

Gene Symbol / ID, Accession #:  TAC1, 6863

RRID:  AB_572266

Reviews

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Reviewed by croninlab
03/02/2017 - 10:27:09 AM
Strong reactivity in crayfish brain

Tissue: Procambarus clarkii, perfused with saline and 4% paraformaldehyde, Brains were embedded in gelatin and sectioned with a vibratome to 60 um.

Protocol: Floating section immunohistochemistry methods followed that of Lin and Strausfeld, 2013.

1.permeabilize tissue with 0.5% PBS-TX
2.wash
3.block in 0.5% PBS-TX + 5% normal goat serum
4.incubate on shaker overnight with Immunostar rabbit anti-Substance P (1:200 in -.5% PBS-TX + 5% NGS)
5.wash
6.incubate on shaker overnight with Life Technologies goat anti-rabbit-Alexa488 (1:400 in 0.5% PBS-TX + 5% NGS)
7.wash
8.mount with Elvanol and coverslip

Results: Clear staining of fibers throughout the brain.

Alice Chou, PhD Candidate
Department of Biology
University of Maryland Baltimore County

Reviewed by user
09/08/2015 - 11:09:17 AM
Success With Adult Zebrafish Brain

We have successfully used the rabbit anti-substance P-antibody (Immunostar #20064) to examine the distribution of substance P in the brain of adult zebrafish.

Zebrafish Brain Preparation:
We perfused zebrafish with PBS and 4% paraformaldehyde and removed the brain after postfixation with 4% paraformaldehyde for two days. Brains were embedded and frozen in OCT and subsequently cut using a Leica cryostat.

Immunofluorescence staining:
Day 1
1. Slides are dried at room temperature for at least 30 min.
2. Wash slides 3X in 1X PBS-T (0.3% Triton X-100).
3. Block slides in 1X PBS-T with 3% normal goat serum for 1 hr at room temperature.
4. Incubate slides with primary in 1X PBS-T with 3% normal goat serum (rabbit anti-substance P, ImmunoStar #20064) overnight at 4 °C.

We store the primary antibody is stored at a 1:10 dilution and used it in a concentration of 1:4,000.

Day 2
5. Remove antiserum and wash tissue sections 3X 5 min with 1X PBS-T.
6. Incubate slides with secondary antibody (e.g. goat anti-rabbit-Alexa488, Life Technologies, diluted 1:500 in PBS-T) for 2 hrs in a wet-chamber covered by tissue to prevent light bleaching of the secondary antibodies.
7. Wash slides 3X 5 min with PBS-T.
8. Mount with Fluoromount (Southern Biotech) and coverslip.

Use fluorescence microscope for visual analysis or imaging.

Thomas Mueller (Ph.D.)
Division of Biology
Kansas State University

Reviewed by droot
07/10/2012 - 10:09:39 PM
Ventral pallidum visualization

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-substance P primary antibody to visualize the entire ventral pallidum.

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-substance P 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 strongly labeled fibers in VP with low background similar to what Haber and Nauta 1983 reported.

David H. Root, Ph.D.
Rutgers University

Reviewed by Dan_Tylee
04/05/2012 - 01:07:22 PM
Strong Staining of Fibers

Tissue: Maccaca fasciscularis, perfused with saline and 4% paraformaldehyde, dehydrated through increase sucrose gradients, sectioned at 40 um.

Method: Immunocytochemistry on free floating sections. Incubated with ImmunoStar's primary antibody at a concentration of 1:5000 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 0.3% Triton X-100. Rinsed, then developed using the instructions for the kit.

Results: Low background. Strong staining of fibers, especially in the pallidum. Clear fiber morphology.

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

Q & A Wall

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we would like to know if the product Substance P cat. # 20064 works on paraffin sections.
Question By Florianna

The Substance P Antibody should be fine in paraffin sections but with appropriate fixation of the tissue and appropriate dilution of the antiserum. Though paraffins will work, they aren't as good as freshly fixed tissue.



Answer By Tech. Support staff on 2015-02-05 12:03:48

we would like to know if the product Substance P cat. # 20064 works on paraffin sections.
Question By robert

The amino acid sequence for ImmunoStar's Substance P Antibody (# 20064) is RPKPQQFFGLM-NH2.

Answer By Colleen on 2013-09-02 12:06:48

MSDS

This product contains the preservative sodium azide. The concentration percent of the sodium azide is ≤ .09%. Although this hazardous substance is a concentration below that required for the preparation of a Material Safety Data Sheet, we created a standard MSDS for your records.

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