Histamine Antibody

$275.00

Product ID: 22939

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Description

Histamine is located in mast cells, endocrine cells of the gut, blood cells and in some cells of the peripheral and central nervous system. Histamine is a potent vasodilator when secreted by mast cells found in various tissues as a result of allergic hypersensitivity or inflammation. In the central nervous system, Histamine is putative neurotransmitter. In the brain, its highest content has been found in the hypothalamus and in certain areas of the mesencephalon. The Histamine antiserum has a sensitivity level capable of detecting the low level Histamine contents of the brain.

The Histamine antiserum was quality control tested using standard immunohistochemical methods. The antiserum demonstrates strongly positive labeling of rat hypothalamus using indirect immunofluorescent and biotin/avidin-HRP techniques. Recommended primary dilution is 1/4000-1/6000 in PBS/0.3% Triton X-100 biotin/avidin-HRP Technique . All staining is blocked by preabsorption of the antiserum with Histamine conjugate. Cross reactivity experiments indicate no cross reactivity with L-histidine or L-histidine containing peptides such as LH-RH.

Photo Description: IHC image of neurons staining for histamine in the rat hypothalamus. The tissue was fixed with 4% carbodimide in 0.05 M phosphate buffered saline, 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. The section was then mounted on slides with permount.

Host: Rabbit

Quantity / Volume:  100 µL

State: Lyophilized Whole Serum

Reacts With: Aplysia Californica (Sea Slug), Bee, Copepods (Crustacean), Crab, Drosophila (Fly), Drosophila Melanogaster (Fly), Flea, Fly, Fruit Fly, Locust, Marine, Monkey, Moth, Mouse, Pig, Pleurobranchaea Japonica (Sea Slug), Rat, Snail

Availability: In Stock

Alternate Names: anti-Histamine

RRID:  AB_572245

Technical Sheets

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.

Download Data Sheet

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Reviews

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Good antibody, mediocre penetration

I have used this antibody (#22939) to label the brains of ~20 different species of insects. Brains are embedded in agarose and sectioned on a vibrating microtome at 50-70um. Note that the 4 hour carbodiimide fixation step (see below) is ABSOLUTELY necessary for the labeling to work. Pre-adsorbing the antibody with KLH does not affect labeling (see Dacks et al 2010, J. Comp. Neurol.). This antibody does not have great penetration for using a vibrating microtome, but 50um sections with 1% TX-100 for incubation with the primary is sufficient.

Day 1:
_____Dissect brain in saline
_____fix brains in 4% 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide in phosphate buffered saline (PBS; pH 6.9) for 4hrs at 4°C
_____Post-fix overnight in 4% paraformaldehyde at 4°C

Day 2:
_____3X 15min PBS washes
_____Embed brains in agarose and section at 50µm
_____4X 15min in PBS with .5% Triton X-100 (PBST) room temperature (RT)
_____Block tissue for 1hr in 2% IgG free Bovine Serum Albumin (BSA)
_____Incubate sections in 2% BSA in PBST (1%TX-100) with 50 mM sodium azide (PBSAT) with 1:500 Histamine (Immunostar # 22939) for 24hrs at RT

Day 3:
_____4 X 15min PBST at RT
_____Block tissue for 1hr (2% BSA)
_____Spin down secondary in PBSAT with 2% BSA for 15min at 14,000RPM
_____Incubate in block (PBSAT) with 1:1000 Cy3 Gt Rb 2° overnight at RT

Day 4:
_____2x 30min wash in PBST
_____2x 30min wash in PBS
_____5 min in 60% glycerol
_____Mount in 80% glycerol

Only use 1% Triton X-100 for primary, otherwise always use .5%.

Andrew Dacks
Assistant Professor
West Virginia University