抗體的生物素化標(biāo)記實驗要點：

1. 血小板衍化生長因子γ抗體品牌 如在反應(yīng)混合液中有疊氮鈉或游離氨基存在,會抑制標(biāo)記反應(yīng)。因此,蛋白質(zhì)在反應(yīng)前要對 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-氨基的密度會有所不同,選擇不當(dāng)則影響標(biāo)記的效率,應(yīng)先用幾個不同的分子比來篩選最適條件；

3.用NHSB量過量也是不利的,抗原的結(jié)合位點可能因此被封閉,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)合位點時,或位于酶的催化位點時,生物素化會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,如羰基、氨基、巰基、異咪唑基及苯酚基,也可與糖基共價結(jié)合；

7.交聯(lián)反應(yīng)后,應(yīng)充分透析,否則,殘余的生物素會對生物素化抗體與親和素的結(jié)合產(chǎn)生競爭作用；

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

產(chǎn)品訂購信息：
英文名稱  Anti-PAFAH1B3/PAFAHG 

中文名稱   血小板衍化生長因子γ抗體品牌 

別    名  PA1B3_HUMAN; PAF acetylhydrolase 29 kDa subunit; PAF AH 29 kDa subunit; PAF AH subunit gamma; PAF-AH 29 kDa subunit; PAF-AH subunit gamma; PAFAH subunit gamma; Pafah1b3; PAFAHG; Platelet activating factor acetylhydrolase IB subunit gamma; Platelet activating factor acetylhydrolase isoform Ib gamma subunit (29kD); Platelet activating factor acetylhydrolase isoform Ib gamma subunit 29kDa; Platelet activating factor acetylhydrolase isoform Ib gamma subunit; Platelet-activating factor acetylhydrolase IB subunit gamma.

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, Horse, Rabbit, Sheep 

產(chǎn)品類型   一抗  

研究領(lǐng)域     心 神經(jīng)生物學(xué) 信號轉(zhuǎn)導(dǎo)

蛋白分子量  predicted molecular weight: 26kDa

性    狀   Lyophilized or Liquid

免 疫 原 KLH conjugated synthetic peptide derived from human PAFAH1B3/PAFAHG 

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

血小板衍化生長因子γ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=1:50-200

（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 The platelet activating factor (PAF) acetylhydrolases catalyze hydrolysis of the sn-2 ester bond of PAF and related pro-inflammatory phospholipids and thus attenuate their bioactivity. The family of PAF acetylhydrolases includes one secreted plasma isozyme and two intracellular isozymes. The intracellular isozymes are distinguished by differences in their primary sequence, tissue localization, subunit composition and substrate preferences. The most thoroughly characterized intracellular isoform, PAFAH1B, is a heterotrimeric protein expressed in brain tissue and plays and important role in brain development and function. PAFAH1B is comprised of a regulatory subunit (LIS1) and two homologous (63% identity) catalytic subunits (PAFAH1B2 and PAFAH1B3), which harbor all the activity of the enzyme. The PAFAH1B2 and PAFAH1B3 subunits readily associate with very high affinity to form heterodimers, and this dimerization is essential for both stability and catalytic activity. PAFAH1B3 is also commonly known as PAFAH1B 29kDa subunit, PAFAH1B subunit or PAFAH1B subunit α1.

Function : Inactivates paf by removing the acetyl group at the sn-2position. This is a catalytic subunit. Plays an important roleduring the development of brain.

Subunit : Cytosolic PAF-AH IB is formed of three subunits of 45 kDa(alpha), 30 kDa (beta) and 29 kDa (gamma). The catalytic activityof the enzyme resides in the beta and gamma subunits, whereas thealpha subunit has regulatory activity. Trimer formation is notessential for the catalytic activity.

Subcellular Location : Cytoplasm.

Similarity : Belongs to the 'GDSL' lipolytic enzyme family.Platelet-activating factor acetylhydrolase IB beta/gamma subunitssubfamily.

Database links : UniProtKB/Swiss-Prot: Q15102.1

抗體的鑒定：

1）血小板衍化生長因子γ抗體品牌 抗體的效價鑒定：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。不同的抗原制備的抗體,要求的效價不一。鑒定效價的方法很多,包括有試管凝集反應(yīng),瓊脂擴散試驗,酶聯(lián)免疫吸附試驗等。常用的抗原所制備的抗體一般都有約成的鑒定效價的方法,以資比較。如制備抗抗體的效價,一般就采用瓊脂擴散試驗來鑒定。

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)的識別能力?？贵w的特異性高,它的識別能力就強。衡量特異性通常以交叉反應(yīng)率來表示。交叉反應(yīng)率可用競爭抑制試驗測定。以不同濃度抗原和近似抗原分別做競爭抑制曲線,計算各自的結(jié)合率,求出各自在IC50時的濃度,并按公式計算交叉反應(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濃度幾乎是無窮大時,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?？贵w親和力的測定對抗體的篩選,確定抗體的用途,驗證抗體的均一性等均有重要意義。

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