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.
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
01/21/2013 - 11:27:32 AM
Good antibody, mediocre penetration
_____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
_____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
_____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
_____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%.
West Virginia University
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Q & A Wall
I am planning to do some single immunoreactions using the biotin streptavidin/HRP technique and I would like to know the right protocol I should follow. I read from the data sheet that the perfusion/fixation should be done
using 4% carbodiimide in PBS. I am interested in doing electron microscopic analysis on this material. Is it possible to add glutaraldehyde(0.1%) to the 4% carbodiimide-PBS?
Question By Icnelia H.
No answer found for this question.
I need to know the cross-reactivity of this antibody.
Question By Katsura
The only cross-reactivity information we have is that which is included on the specification sheet: no cross-reactivity with L-histidine or L-histidine-containing peptides such as LH-RH. This testing was done in-house, by dot-blot.
Answer By Jodi on 2009-03-06 12:02:25
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 MSDS