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ELISA试剂盒增食欲素A/欲激素A抗体操作步骤
更新时间:2014-04-28      阅读:1466

ELISA试剂盒增食欲素A/欲激素A抗体英文名称  Anti-Orexin A 
中文名称  增食欲素A/欲激素A抗体 
别    名  orexin-A; HCRT 1; HCRT; HCRT1; Hypocretin (orexin) neuropeptide precursor; Hypocretin 1; Hypocretin A; Hypocretin; Hypocretin neuropeptide precursor; Hypocretin1; Orexin A; Orexin; Orexin precursor; OrexinA; OX; PPORX; PPOX; PREPROOREXIN; ORX-A; OREX_HUMAN. 
浓    度  1mg/1ml 
规 格  0.2ml/200μg
抗体来源  Rabbit  
克隆类型  polyclonal 
交叉反应  Human, Mouse, Rat, Dog, Pig, Cow, Sheep  
产品类型  一抗    
研究领域  生长因子和激素 内分泌病  
蛋白分子量  predicted molecular weight: 3.6kDa 
性    状  Lyophilized or Liquid 
免 疫 原  KLH conjugated synthetic peptide derived from human Orexin A 
亚    型  IgG 
纯化方法  affinity purified by Protein A 
储 存 液  Preservative: 15mM Sodium Azide, Constituents: 1% BSA, 0.01M PBS, pH 7.4 
ELISA试剂盒增食欲素A/欲激素A抗体产品应用   WB=1:100-500  ELISA=1:500-1000  IP=1:20-100  IHC-P=1:100-500  IHC-F=1:100-500  ICC=1:100-500  IF=1:100-500 
(石蜡切片需做抗原修复) 
 not yet tested in other applications.
 optimal dilutions/concentrations should be determined by the end user.  
保存条件  Store at -20 °C for one year. Avoid repeated freeze/thaw cycles. The lyophilized antibody is stable at room temperature for at least one month and for greater than a year when kept at -20°C. When reconstituted in sterile pH 7.4 0.01M PBS or diluent of antibody the antibody is stable for at least two weeks at 2-4 °C. 
Important Note  This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications. 
产品介绍 This gene encodes a hypothalamic neuropeptide precursor protein that gives rise to two mature neuropeptides, orexin A and orexin B, by proteolytic processing. Orexin A and orexin B, which bind to orphan G-protein coupled receptors HCRTR1 and HCRTR2, function in the regulation of sleep and arousal. This neuropeptide arrangement may also play a role in feeding behavior, metabolism, and homeostasis.
Function : Neuropeptides that play a significant role in the regulation of food intake and sleep-wakefulness, possibly by coordinating the complex behavioral and physiologic responses of these complementary homeostatic functions. A broader role in the homeostatic regulation of energy metabolism, autonomic function, hormonal balance and the regulation of body fluids, is also suggested. Orexin-A binds to both OX1R and OX2R with a high affinity, whereas orexin-B binds only to OX2R with a similar high affinity. 
Subcellular Location : Rough endoplasmic reticulum (By similarity). Cytoplasmic vesicle (By similarity). Cell junction, synapse (By similarity). Note=Associated with perikaryal rough endoplasmic reticulum as well as cytoplasmic large granular vesicles at synapses (By similarity). 
Tissue Specificity : Abundantly expressed in subthalamic nucleus but undetectable in other brain regions tested (hypothalamus was not tested) and in heart, placenta, lung, liver, skeletal muscle, kidney and pancreas.
Post-translational modifications : Specific enzymatic cleavages at paired basic residues yield the different active peptides. 
DISEASE : Defects in HCRT are the cause of narcolepsy type 1 (NRCLP1) [MIM:161400]. Narcolepsy is a neurological disabling sleep disorder, characterized by excessive daytime sleepiness, sleep fragmentation, symptoms of abnormal rapid-eye-movement (REM) sleep, such as cataplexy, hypnagogic hallucinations, and sleep paralysis. Cataplexy is a sudden loss of muscle tone triggered by emotions, which is the most valuable clinical feature used to diagnose narcolepsy. Human narcolepsy is primarily a sporadically occurring disorder but familial clustering has been observed. Note=Human narcolepsy is associated with a deficient orexin system. Orexins are absent and/or greatly diminished in the brain and cerebrospinal fluid (CSF) of most narcoleptic patients. 
Similarity : Belongs to the orexin family. 
Database links : UniProtKB/Swiss-Prot: O43612.1
ELISA试剂盒操作步骤
使用前,将所有试剂充分混匀。不要使液体产生大量的泡沫,以免加样时加入大量的气泡,产生加样上的误差。
根据待测样品数量加上标准品的数量决定所需的板条数。每个标准品和空白孔建议做复孔。每个样品根据自己的数量来定,能使用复孔的尽量做复孔。标本用标本稀释液1:1稀释后加入50ul于反应孔内。
加入稀释好后的标准品50ul于反应孔、加入待测样品50ul于反应孔内。立即加入50ul的生物素标记的抗体。盖上膜板,轻轻振荡混匀,37℃温育1小时。
甩去孔内液体,每孔加满洗涤液,振荡30秒,甩去洗涤液,用吸水纸拍干。重复此操作3次。如果用洗板机洗涤,洗涤次数增加一次。
每孔加入80ul的亲和链酶素-HRP,轻轻振荡混匀,37℃温育30分钟。
甩去孔内液体,每孔加满洗涤液,振荡30秒,甩去洗涤液,用吸水纸拍干。重复此操作3次。如果用洗板机洗涤,洗涤次数增加一次。
每孔加入底物A、B各50ul,轻轻振荡混匀,37℃温育10分钟。避免光照。
取出酶标板,迅速加入50ul终止液,加入终止液后应立即测定结果。
在450nm波长处测定各孔的OD值。

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