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 格  0.2ml/200μg
抗體來源  Rabbit  
克隆類型  polyclonal 
交叉反應  Human, Mouse, Rat, Dog, Pig, Cow, Sheep  
產品類型  一抗    
研究領域  生長因子和激素 內分泌病  
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 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抗體產品應用   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） 
 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。不要使液體產生大量的泡沫，以免加樣時加入大量的氣泡，產生加樣上的誤差。
根據待測樣品數量加上標準品的數量決定所需的板條數。每個標準品和空白孔建議做復孔。每個樣品根據自己的數量來定，能使用復孔的盡量做復孔。標本用標本稀釋液1：1稀釋后加入50ul于反應孔內。
加入稀釋好后的標準品50ul于反應孔、加入待測樣品50ul于反應孔內。立即加入50ul的生物素標記的抗體。蓋上膜板，輕輕振蕩混勻，37℃溫育1小時。
甩去孔內液體，每孔加滿洗滌液，振蕩30秒，甩去洗滌液，用吸水紙拍干。重復此操作3次。如果用洗板機洗滌，洗滌次數增加一次。
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。重復此操作3次。如果用洗板機洗滌，洗滌次數增加一次。
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。
取出酶標板，迅速加入50ul終止液，加入終止液后應立即測定結果。
在450nm波長處測定各孔的OD值。