抗體的生物素化標記實驗要點：

1. 蛋白磷酸酯酶-1B抗體費用 如在反應混合液中有疊氮鈉或游離氨基存在,會抑制標記反應。因此,蛋白質(zhì)在反應前要對 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,選擇不當則影響標記的效率,應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,抗原的結合位點可能因此被封閉,導致抗體失活；

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.當游離ε-氨基(賴氨酸殘基的氨基)存在于抗體的抗原結合位點時,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,也可與糖基共價結合；

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

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

產(chǎn)品訂購信息：
英文名稱  Anti-PP-1B 

中文名稱   蛋白磷酸酯酶-1B抗體費用 

別    名  Protein phosphates-1; PP-1 gamma, beta, alpha, catalytic subunit; MGC3672; PP 1B; PP-1B; PPP1CB; PP1B; PPP1CD; PP1beta; Protein phosphatase 1 beta; Protein phosphatase 1 catalytic subunit beta isoform; Protein phosphatase 1 delta; Serine threonine protein phosphatase PP1 beta catalytic subunit; Serine/threonine-protein phosphatase PP1-beta catalytic subunit; PP1B_HUMAN; Protein phosphatase 1, catalytic subunit, beta isozyme; Protein phosphatase 1, catalytic subunit, delta isoform;

濃    度   1mg/1ml

規(guī) 格  0.1ml/100μg 0.2ml/200μg

抗體來源   Mouse

克隆類型   polyclonal

交叉反應   Human, Mouse, Rat, Cow, Horse, Rabbit 

產(chǎn)品類型   一抗  

研究領域    細胞生物 神經(jīng)生物學 信號轉(zhuǎn)導 細胞凋亡 激酶和磷酸酶

蛋白分子量  predicted molecular weight: 37kDa

性    狀   Lyophilized or Liquid

免 疫 原 KLH conjugated synthetic peptide derived from human PP-1B C-terminus 

亞    型   IgG

純化方法   affinity purified by Protein A

儲 存 液   0.01M PBS, pH 7.4 with 10 mg/ml BSA and 0.1% Sodium azide

蛋白磷酸酯酶-1B抗體費用 產(chǎn)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 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(chǎn)品介紹 The phosphorylation and de phosphorylation of proteins on serine and threonine residues is an essential means of regulating a broad range of cellular functions in eukaryotes, including division, homeostasis and apoptosis. A geoup of proteins that are intimately involved in this process are the serine/ threonine protein phosphatases. The protein phosphatases (PP) holoenzyme is a trimertic complex composed of a regulatory subunit, a variable subunit and a catalytic subunit. Four major families of protein phosphatase catalytic subunits have been identified and are designated PP1, PP2A, PP2B (calcineurin) and PP2C. An additional protein phosphatase catalytic subunit, PPX or PP4, is the member of a potentially novel PP family. The PP1 family is composed of subfamily members PP1 alpha, PP1 beta,and PP1 gamma.The PP2A family iscomposed of subfamily members PP2A alpha,and PP2A beta. The PP2B family is composed of subfamily members PP2B-A alpha, PP2B-A beta and PP2B-A gamma. The PP2C family is composed of PP2C alpha, PP2C beta, PP2C gamma. Wip1, a protein isentified in the p53 DNA response pathway, has also been identified as a potential member of the PP2C family. Regulatary subunits include dnuclear inhibitor of PP1(NIPP1), PP2A-AAlphaand–A beta, PP2A-B alpha and B beta,PP2A-C alpha and C beta, PP2A-B56-alpha and–B56 beta, PR48 and PP2B-B1 and–B2.

Function : Protein phosphatase that associates with over 200 regulatory proteins to form highly specific holoenzymes which dephosphorylate hundreds of biological targets. Protein phosphatase (PP1) is essential for cell division, it participates in the regulation of glycogen metabolism, muscle contractility and protein synthesis. Involved in regulation of ionic conductances and long-term synaptic plasticity. Component of the PTW/PP1 phosphatase complex, which plays a role in the control of chromatin structure and cell cycle progression during the transition from mitosis into interphase.

Subunit : PP1 comprises a catalytic subunit, PPP1CA, PPP1CB or PPP1CC, which is folded into its native form by inhibitor 2 and glycogen synthetase kinase 3, and then complexed to one or several targeting or regulatory subunits. The targeting or regulatory subunits determine the substrate specificity of PP1. PPP1R12A, PPP1R12B and PPP1R12C mediate binding to myosin. PPP1R3A (in skeletal muscle), PPP1R3B (in liver), PPP1R3C, PPP1R3D and PPP1R3F (in brain) mediate binding to glycogen. Part of a complex containing PPP1R15B, PP1 and NCK1/2 (By similarity). Component of the MLL5-L complex, at least composed of MLL5, STK38, PPP1CA, PPP1CB, PPP1CC, HCFC1, ACTB and OGT. Interacts with PPP1R7 and PPP1R12C. PPP1R15A and PPP1R15B mediate binding to EIF2S1. Interacts with PPP1R16B. Component of the PTW/PP1 phosphatase complex, composed of PPP1R10/PNUTS, TOX4, WDR82, and PPP1CA or PPP1CB or PPP1CC. Interacts with PPP1R8. Interacts with TRIM28; the interaction is weak. Interacts with PPP1R12A and NUAK1; the interaction is direct.

Subcellular Location : Cytoplasm. Nucleus. Nucleus, nucleoplasm. Nucleus, nucleolus. Note=Highly mobile in cells and can be relocalized through interaction with targeting subunits. In the presence of PPP1R8 relocalizes from the nucleus to nuclear speckles.

Database links : UniProtKB/Swiss-Prot: P62140.3

抗體的鑒定：

1）蛋白磷酸酯酶-1B抗體費用 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,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率來表示。交叉反應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,計算各自的結合率,求出各自在IC50時的濃度,并按公式計算交叉反應率。 

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,抗體分子的結合位點與抗原決定簇之間立體構型的合適度決定的。有助于維持抗原抗體復合物穩(wěn)定的分子間力有氫鍵,疏水鍵,側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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Adipo R1(adiponectin receptor 1) 脂聯(lián)素受體-1(抗原) 0.5mg

IBP160 英文名稱： 160kDa內(nèi)含子結合蛋白抗體 0.2ml

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Anti-HOXc8/FITC 熒光素標記同源盒基因的一種抗體IgGMulti-class antibodies規(guī)格： 0.2ml

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Anti-phospho-Smad2(p-Ser465) /FITC 熒光素標記兔抗人、大、小鼠、，馬，犬，雞磷酸化Smad2抗體IgGMulti-class antibodies規(guī)格： 0.2ml

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Rabbit Anti-human IgG/Alexa Fluor 488 Alexa Fluor 488標記的兔抗人IgG 0.1ml

GLYATL1 英文名稱： 甘胺酸-N-?；D(zhuǎn)移酶樣1抗體 0.2ml

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IDUA(Mouse Alpha-L-iduronidase) ELISA Kit 小鼠αL艾杜糖苷酸酶 96T

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犬腎細胞系/IgR;MDCK/IgR

PLAT Others Human 人 PLAT / tPA 人細胞裂解液 (陽性對照)

蛋白磷酸酯酶-1B抗體費用 CD5 Others Rat 大鼠 CD5 人細胞裂解液 (陽性對照)

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