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    您當(dāng)前的位置:首頁 > 技術(shù)文章 > ELISA試劑盒增食欲素A/欲激素A抗體操作步驟

    ELISA試劑盒增食欲素A/欲激素A抗體操作步驟

    發(fā)布時(shí)間: 2014-04-28  點(diǎn)擊次數(shù): 812次

    ELISA試劑盒增食欲素A/欲激素A抗體英文名稱  Anti-Orexin A 
    中文名稱  增食欲素A/欲激素A抗體 
    別    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. 
    濃    度  1mg/1ml 
    規(guī) 格  0.2ml/200μg
    抗體來源  Rabbit  
    克隆類型  polyclonal 
    交叉反應(yīng)  Human, Mouse, Rat, Dog, Pig, Cow, Sheep  
    產(chǎn)品類型  一抗    
    研究領(lǐng)域  生長因子和激素 內(nèi)分泌病  
    蛋白分子量  predicted molecular weight: 3.6kDa 
    性    狀  Lyophilized or Liquid 
    免 疫 原  KLH conjugated synthetic peptide derived from human Orexin A 
    亞    型  IgG 
    純化方法  affinity purified by Protein A 
    儲(chǔ) 存 液  Preservative: 15mM Sodium Azide, Constituents: 1% BSA, 0.01M PBS, pH 7.4 
    ELISA試劑盒增食欲素A/欲激素A抗體產(chǎn)品應(yīng)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 
    (石蠟切片需做抗原修復(fù)) 
     not yet tested in other applications.
     optimal dilutions/concentrations should be determined by the end user.  
    保存條件  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. 
    產(chǎn)品介紹 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試劑盒操作步驟
    使用前,將所有試劑充分混勻。不要使液體產(chǎn)生大量的泡沫,以免加樣時(shí)加入大量的氣泡,產(chǎn)生加樣上的誤差。
    根據(jù)待測(cè)樣品數(shù)量加上標(biāo)準(zhǔn)品的數(shù)量決定所需的板條數(shù)。每個(gè)標(biāo)準(zhǔn)品和空白孔建議做復(fù)孔。每個(gè)樣品根據(jù)自己的數(shù)量來定,能使用復(fù)孔的盡量做復(fù)孔。標(biāo)本用標(biāo)本稀釋液1:1稀釋后加入50ul于反應(yīng)孔內(nèi)。
    加入稀釋好后的標(biāo)準(zhǔn)品50ul于反應(yīng)孔、加入待測(cè)樣品50ul于反應(yīng)孔內(nèi)。立即加入50ul的生物素標(biāo)記的抗體。蓋上膜板,輕輕振蕩混勻,37℃溫育1小時(shí)。
    甩去孔內(nèi)液體,每孔加滿洗滌液,振蕩30秒,甩去洗滌液,用吸水紙拍干。重復(fù)此操作3次。如果用洗板機(jī)洗滌,洗滌次數(shù)增加一次。
    每孔加入80ul的親和鏈酶素-HRP,輕輕振蕩混勻,37℃溫育30分鐘。
    甩去孔內(nèi)液體,每孔加滿洗滌液,振蕩30秒,甩去洗滌液,用吸水紙拍干。重復(fù)此操作3次。如果用洗板機(jī)洗滌,洗滌次數(shù)增加一次。
    每孔加入底物A、B各50ul,輕輕振蕩混勻,37℃溫育10分鐘。避免光照。
    取出酶標(biāo)板,迅速加入50ul終止液,加入終止液后應(yīng)立即測(cè)定結(jié)果。
    在450nm波長處測(cè)定各孔的OD值。

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