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產(chǎn)地 | 進(jìn)口、國產(chǎn) |
品牌 | 上海莼試 |
保存條件 | Store at -20 °C |
貨號 | CS10789 |
應(yīng)用范圍 | 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 |
CAS編號 | |
抗體名 | Anti-PPIA/Cyclophilin A/CYPA |
克隆性 | |
靶點(diǎn) | 詳見說明書 |
適應(yīng)物種 | 詳見說明書 |
形態(tài) | 詳見說明書 |
宿主 | 詳見說明書 |
亞型 | IgG |
標(biāo)識物 | 詳見說明書 |
濃度 | 1mg/1ml% |
免疫原 | KLH conjugated synthetic peptide derived from human Cyclophilin A. |
抗體的生物素化標(biāo)記實(shí)驗(yàn)要點(diǎn):
1. 親環(huán)蛋白(親環(huán)素)PPIA抗體費(fèi)用 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 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(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(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,故對某些細(xì)胞可導(dǎo)致高本底。
產(chǎn)品訂購信息:
英文名稱 Anti-PPIA/Cyclophilin A/CYPA
中文名稱 親環(huán)蛋白(親環(huán)素)PPIA抗體費(fèi)用
別 名 Cyclophilin A; CyclophilinA; Cyclosporin A binding protein; Cyclosporin A-binding protein; CYPA; CYPH; Peptidyl prolyl cis trans isomerase A; Peptidyl-prolyl cis-trans isomerase A; Peptidylprolyl isomerase A; PPIA; PPIA protein; PPIA_HUMAN; PPIase A; Rotamase A; RotamaseA; T cell cyclophilin.
濃 度 1mg/1ml
規(guī) 格 0.1ml/100μg 0.2ml/200μg
抗體來源 Rabbit
克隆類型 polyclonal
交叉反應(yīng) Human, Mouse, Rat, Dog, Pig, Cow, Rabbit
產(chǎn)品類型 一抗
研究領(lǐng)域 免疫學(xué) 信號轉(zhuǎn)導(dǎo) t-淋巴細(xì)胞
蛋白分子量 predicted molecular weight: 18kDa
性 狀 Lyophilized or Liquid
免 疫 原 KLH conjugated synthetic peptide derived from human Cyclophilin A.
亞 型 IgG
純化方法 affinity purified by Protein A
儲(chǔ) 存 液 0.01M PBS, pH 7.4 with 10 mg/ml BSA and 0.1% Sodium azide
親環(huán)蛋白(親環(huán)素)PPIA抗體費(fèi)用 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 The immunosuppressant cyclosporin A (CsA) forms a trimolecular complex with cyclophilin and calcineurins to inhibit calcineurin phosphatase activity (1). Cyclophilins are conserved, ubiquitous and abundant cytosolic peptidyl-prolyl cis-trans isomerases that accelerate the isomerization of XaaPro peptide bonds and the refolding of proteins (2,3). Human cyclophilin A (CyPA), an intracellular protein of 165 amino acids, is the target of the CsA and is encoded by a single unique gene conserved from yeast to humans (4,5). CyPA is known for its involvement in T cell differentiation and proliferation and is highly expressed in brain (6). CyPA is incorporated into the virion of the type 1 human immunodeficiency virus (HIV-1) via a direct interaction with the capsid domain of the viral Gag polyprotein and is crucial for efficient viral replication (7,8). Cyclophilin B (CyPB) is a member of the cyclophilin family with specific N- and C-terminal extensions. Unlike CyPA, CyPB has a signal sequence leading to its translocation in the endoplasmic reticulum. CyPB is secreted in biological fluids such as blood or milk and binds to a specific receptor present on the human lymphoblastic cell line Jurkat and on human peripheral blood lymphocytes (9,10).
Function : PPIases accelerate the folding of proteins. It catalyzes the cis-trans isomerization of proline imidic peptide bonds in oligopeptides.
Subcellular Location : Cytoplasm.
Similarity : Belongs to the cyclophilin-type PPIase family. PPIase A subfamily.
抗體的鑒定:
1)親環(huán)蛋白(親環(huán)素)PPIA抗體費(fèi)用 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(qiáng)。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(yàn)測定。以不同濃度抗原和近似抗原分別做競爭抑制曲線,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(yàn)證抗體的均一性等均有重要意義。
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