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品种反应性 主要应用 宿主 格式 抗体类型 R IHC, ICC Rb Serum Polyclonal Antibody 描述 产品目录编号 AB5885 Replaces AB134 品牌系列 Chemicon®
商名 - Chemicon
描述 Anti-Histamine Antibody 产品信息 格式 Serum 演示 Rabbit antisera. Liquid containing 0.1% sodium azide. 应用 应用 Anti-Histamine Antibody is an antibody against Histamine for use in IC, IH. 主要应用 - Immunohistochemistry
- Immunocytochemistry
应用说明 Immunohistoschemistry on brain and stomach using PAP or ABC: 1:1,000-1:4,000. Suggested fixative is 4% EDAC (1-ethyl-3(3-dimethylaminoproply)-carbodiimide) or EDAC + PFA.
Immunocytochemistry on cultured cells (2H3 mastocytoma): 1:100-1:2,000. This antibody is not suitable for use on samples fixed only with aldehydes.
Optimal working dilutions must be determined by end user.
SUGGESTED IMMUNOCHEMISTRY PROTOCOLS FOR AB5885
Immunohistochemistry for vertebrates
The histamine antibody can be applied to different species and tissues. The serum gives good results only when used on properly fixed tissue sections. Specific detection of histamine-like immunoreactivity requires the use of EDAC (1-ethyl-3(3-dimethylaminoproply)-carbodiimide) fixative. Immersion fixation is suitable for small animals and tissue blocks. Perfusion is recommended for vertebrates.
Solutions
1. 0.1 M sodium phosphate buffer, pH 7.4
2. 4% EDAC (1-ethyl-3(3-dimethylaminoproply)-carbodiimide) in 0.1M sodium phosphate buffer (pH 7.4);
the final pH will be around 7.0. Always use freshly prepared solution.
3. 0.9% saline
Application to small laboratory animals (vertebrates)
1. Open the thorax under suitable deep anesthesia (e.g. pentobarbital). Open the right heart auricle with small scissors, Perfuse through the left ventricle or ascending aorta with saline until all blood is washed out (about 50 mL in a rat).
2. Perfuse with fresh 4% EDAC through the same needle. Use about 1 mL/gram of body weight.
3. Remove the tissues and immerse them in 4% EDAC for 2 hours or overnight.
4. Wash the tissue in 0.1 M sodium phosphate buffer containing 20-30% sucrose overnight.
5. Freeze the tissues and make cryostat sections for immunohistochemistry.
Tissues from many species can be fixed by immersion in 4% EDAC overnight at 2-8°C. The pieces should be no thicker than 0.5 cm because the fixative does not pentrate very well. Note that the tissues are not firm hard after fixation with EDAC. Handling free floating sections requires experience, sections collected on adhesive-coated slides are easy to handle. If aldehyde fixation is necessary for e.g double staining procedures, EDAC perfusion should be followed by perfusion with 4% paraformaldehyde in the same buffer (and if needed 0.5% glutaraldehyde + 0.2% saturated picric acid). However these samples must be treated with 0.5% borohydride after sectioning. Incubation with the histamine antiserum (diluted in PBS with 0.25% Triton) is best performed at 2-8°C for 24-48 hours. Extended incubation times of up to 5 days have also been used successfully.
Immunocytochemistry for cell culture
The histamine antibody can be applied to primary cell cultures and cell lines from different species and tissues. The serum gives good results only when used on properly fixed tissue sections. Specific detection of histaminelike immunoreactivity requires the use of EDAC (1-ethyl-3(3-dimethylaminoproply)-carbodiimide) fixative. Immersion fixation is suitable for small animals and tissue blocks. Perfusion is recommended for vertebrates.
Solutions
1. 0.1 M sodium phosphate buffer, pH 7.4
2. 4% EDAC (1-ethyl-3(3-dimethylaminoproply)-carbodiimide) in 0.1M sodium phosphate buffer (pH 7.4); the final pH will be around 7.0. Always use freshly prepared solution.
3. 0.9% saline
Incubation procedure
1. Pour out the culture medium and wash the cells with 0.9% saline or 0.1 M sodium phosphate buffer. (This can be omitted if the cells are sensitive to changes.)
2. Pour in freshly prepared 4% EDAC, fix at 2-8°C for 0.5-12 hours depending on the thickness of the culture.
3. Wash with 0.1 M sodium phosphate buffer or PBS 2 x 10 min.
4. Preincubate with normal serum from the species used for production of the secondary antiserum for 20 minutes at room temperature. Pour out the normal serum, do not wash.
5. Incubate with the histamine antiserum diluted appropriately (1:200-1:5,000) for 24-48 hours at 2-8°C.
If electron microscopy or double staining procedures with other antibodies are needed, the cells can be fixed after 4% EDAC with a mixture of 4% paraformaldehyde, 0.5% glutaraldehyde and 0.2% saturated picric acid for 30 minutes. After thorough washing, these samples must be treated with 0.5% sodium borohydride (and washed
extensively) before incubation.
Any routinely used detection method (indirect immunofluorescence, PAP procedure, biotin-streptavidin peroxidase) can be used.生物信息 免疫原品种 Histamine conjugated to KLH with EDAC. 宿主 Rabbit 特异性 Histamine. No reactivity to L-histadine containing peptides. Preabsorption of the antiserum with a histamine-BSA or histamine-ovalbumin conjugate (1-100 mg/mL depending on tissue) completely abolished the specific reaction.
SPECIES REACTIVITIES:
Vertebrates and invertebrates (human, rat, mouse, frog, fish, mollusks, flatworm).品种反应性 Rat 抗体类型 Polyclonal Antibody 产品使用声明 使用声明 - Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
储存和货运信息 存储条件 Maintain at -20°C in undiluted aliquots for up to 6 months after date of receipt. Avoid repeated freeze/thaw cycles. 包装信息 数量 100 µL
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