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抗體的生物素化標(biāo)記實(shí)驗(yàn)要點(diǎn)：

1. 過(guò)氧化物酶體增殖物激活受體γ輔激活子1β抗體品牌 如在反應(yīng)混合液中有疊氮鈉或游離氨基存在,會(huì)抑制標(biāo)記反應(yīng)。因此,蛋白質(zhì)在反應(yīng)前要對(duì) 0.1mol/L碳酸氫鈉緩沖液或0.5mol/L硼酸緩沖液充分透析；

2.所用的NHSB及待生物素化蛋白質(zhì)之間的分子比按蛋白質(zhì)表面的ε-氨基的密度會(huì)有所不同,選擇不當(dāng)則影響標(biāo)記的效率,應(yīng)先用幾個(gè)不同的分子比來(lái)篩選最適條件；

3.用NHSB量過(guò)量也是不利的,抗原的結(jié)合位點(diǎn)可能因此被封閉,導(dǎo)致抗體失活；

4.由于抗體的氨基不易接近可能造成生物素化不足,此時(shí)可加入去污劑如 Triton x-100, Tween20等；

5.當(dāng)游離ε-氨基(賴氨酸殘基的氨基)存在于抗體的抗原結(jié)合位點(diǎn)時(shí),或位于酶的催化位點(diǎn)時(shí),生物素化會(huì)降低或損傷抗體蛋白的結(jié)合力或活性；

6.生物素還可能與不同的功能基團(tuán),如羰基、氨基、巰基、異咪唑基及苯酚基,也可與糖基共價(jià)結(jié)合；

7.交聯(lián)反應(yīng)后,應(yīng)充分透析,否則,殘余的生物素會(huì)對(duì)生物素化抗體與親和素的結(jié)合產(chǎn)生競(jìng)爭(zhēng)作用；

8.在細(xì)胞的熒光標(biāo)記實(shí)驗(yàn)中,中和親和素的本底低,但由于鏈霉親和素含有少量正電荷,故對(duì)某些細(xì)胞可導(dǎo)致高本底。

產(chǎn)品訂購(gòu)信息：
英文名稱  Anti-PGC1 beta 

中文名稱   過(guò)氧化物酶體增殖物激活受體γ輔激活子1β抗體品牌 

別    名  PERC; peroxisome proliferative activated receptor, gamma, coactivator 1,; peroxisome proliferator-activated receptor gamma coactivator 1 beta; Peroxisome proliferator-activated receptor gamma coactivator 1-beta; peroxisome proliferator-activated receptor gamma, coactivator 1 beta; PGC-1(beta); PGC-1-beta; PGC-1-related estrogen receptor alpha coactivator; PGC1; PPAR gamma coactivator-1beta; PPAR-gamma coactivator 1-beta; PPARGC-1-beta; PPARGC1; Ppargc1b; PRGC2_HUMAN.

濃    度   1mg/1ml

規(guī) 格  0.2ml/200μg

抗體來(lái)源   Rabbit

克隆類型   polyclonal

交叉反應(yīng)   Human, Mouse, Rat, Dog, Pig, Cow, Rabbit, Guinea Pig 

產(chǎn)品類型   一抗  

研究領(lǐng)域    心 細(xì)胞生物 信號(hào)轉(zhuǎn)導(dǎo) 表觀遺傳學(xué)

蛋白分子量  predicted molecular weight: 113kDa

性    狀   Lyophilized or Liquid

免 疫 原 KLH conjugated synthetic peptide derived from human PGC1 beta 

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

過(guò)氧化物酶體增殖物激活受體γ輔激活子1β抗體品牌 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 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 Plays a role of stimulator of transcription factors and nuclear receptors activities. Activates transcritional activity of estrogen receptor alpha, nuclear respiratory factor 1 (NRF1) and glucocorticoid receptor in the presence of glucocorticoids. May play a role in constitutive non-adrenergic-mediated mitochondrial biogenesis as suggested by increased basal oxygen consumption and mitochondrial number when overexpressed. May be involved in fat oxidation and non-oxidative glucose metabolism and in the regulation of energy expenditure.

Tissue specificity: Ubiquitous with higher expression in heart, brain and skeletal muscle.

Function : Plays a role of stimulator of transcription factors andnuclear receptors activities. Activates transcritional activity ofestrogen receptor alpha, nuclear respiratory factor 1 (NRF1) andglucocorticoid receptor in the presence of glucocorticoids. Mayplay a role in constitutive non-adrenergic-mediated mitochondrialbiogenesis as suggested by increased basal oxygen consumption andmitochondrial number when overexpressed. May be involved in fatoxidation and non-oxidative glucose metabolism and in theregulation of energy expenditure.

Subunit : Interacts with hepatocyte nuclear factor 4-alpha/HNF4A,Sterol regulatory binding transcription factor 1/SREBF1,PPAR-alpha/PPARA, thyroid hormone receptor beta/THRB and host cellfactor/HCFC1. Interacts with estrogen-related receptor gamma/ESRRGand alpha/ESRRA. Interacts with PRDM16 (By similarity). Interactswith estrogen receptor alpha/ESR1.

Tissue Specificity : Ubiquitous with higher expression in heart,brain and skeletal muscle.

Similarity : Contains 1 RRM (RNA recognition motif) domain.

Database links :

UniProtKB/Swiss-Prot: Q86YN6.2

Entrez Gene: 133522 Human

Entrez Gene: 170826 Mouse

Entrez Gene: 291567 Rat

Omim: 608886 Human

SwissProt: Q86YN6 Human

SwissProt: Q8VHJ7 Mouse

SwissProt: Q811R2 Rat

Unigene: 483816 Human

抗體的鑒定：

1）過(guò)氧化物酶體增殖物激活受體γ輔激活子1β抗體品牌 抗體的效價(jià)鑒定：不管是用于診斷還是用于,制備抗體的目的都是要求較高效價(jià)。不同的抗原制備的抗體,要求的效價(jià)不一。鑒定效價(jià)的方法很多,包括有試管凝集反應(yīng),瓊脂擴(kuò)散試驗(yàn),酶聯(lián)免疫吸附試驗(yàn)等。常用的抗原所制備的抗體一般都有約成的鑒定效價(jià)的方法,以資比較。如制備抗抗體的效價(jià),一般就采用瓊脂擴(kuò)散試驗(yàn)來(lái)鑒定。

2）抗體的特異性鑒定：抗體的特異性是指與相應(yīng)抗原或近似抗原物質(zhì)的識(shí)別能力。抗體的特異性高,它的識(shí)別能力就強(qiáng)。衡量特異性通常以交叉反應(yīng)率來(lái)表示。交叉反應(yīng)率可用競(jìng)爭(zhēng)抑制試驗(yàn)測(cè)定。以不同濃度抗原和近似抗原分別做競(jìng)爭(zhēng)抑制曲線,計(jì)算各自的結(jié)合率,求出各自在IC50時(shí)的濃度,并按公式計(jì)算交叉反應(yīng)率。 

如果所用抗原濃度IC50濃度為pg/管,而一些近似抗原物質(zhì)的IC50濃度幾乎是無(wú)窮大時(shí),表示這一抗血清與其他抗原物質(zhì)的交叉反應(yīng)率近似為0,即該血清的特異性較好。

3）抗體親和力：是指抗體和抗原結(jié)合的牢固程度。親和力的高低是由抗原分子的大小,抗體分子的結(jié)合位點(diǎn)與抗原決定簇之間立體構(gòu)型的合適度決定的。有助于維持抗原抗體復(fù)合物穩(wěn)定的分子間力有氫鍵,疏水鍵,側(cè)鏈相反電荷基因的庫(kù)侖力,范德華力和空間斥力。親和力常以親和常數(shù)K表示,K的單位是L/mol。抗體親和力的測(cè)定對(duì)抗體的篩選,確定抗體的用途,驗(yàn)證抗體的均一性等均有重要意義。

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