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產(chǎn)地 | 進口、國產(chǎn) |
品牌 | 上海莼試 |
保存條件 | Store at -20 °C |
貨號 | CS10356 |
應用范圍 | WB=1:100-500 ELISA=1:500-1000 IHC-P=1:100-500 IHC-F=1:100-500 ICC=1:100-500 IF=1:100-500 |
CAS編號 | |
抗體名 | Anti-PNUTS/PPP1R10 |
克隆性 | |
靶點 | 詳見說明書 |
適應物種 | 詳見說明書 |
形態(tài) | 詳見說明書 |
宿主 | 詳見說明書 |
亞型 | IgG |
標識物 | 詳見說明書 |
濃度 | 1mg/1ml% |
免疫原 | KLH conjugated synthetic peptide derived from human PNUTS/PPP1R10 |
抗體的生物素化標記實驗要點:
1. 絲氨酸/蘇氨酸蛋白磷酸酶1調(diào)節(jié)亞基10抗體說明書 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(jié)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(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(jié)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-PNUTS/PPP1R10
中文名稱 絲氨酸/蘇氨酸蛋白磷酸酶1調(diào)節(jié)亞基10抗體說明書
別 名 CAT 53; CAT53; FB 19; FB19; FB19 protein; MHC class I region proline rich protein CAT53; p99; Phosphatase 1 nuclear targeting subunit; Phosphatase nuclear targeting subunit; PNUTS; PP1 binding protein of 114 kDa; PP1R10; Protein phosphatase 1 regulatory (inhibitor) subunit 10; Protein phosphatase 1 regulatory subunit 10; Serine/threonine protein phosphatase 1 regulatory subunit 10; PP1RA_HUMAN.
濃 度 1mg/1ml
規(guī) 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(chǎn)品類型 一抗
研究領(lǐng)域 神經(jīng)生物學 信號轉(zhuǎn)導 激酶和磷酸酶 Alzheimer's
蛋白分子量 predicted molecular weight: 99kDa
性 狀 Lyophilized or Liquid
免 疫 原 KLH conjugated synthetic peptide derived from human PNUTS/PPP1R10
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
絲氨酸/蘇氨酸蛋白磷酸酶1調(diào)節(jié)亞基10抗體說明書 產(chǎn)品應用 WB=1:100-500 ELISA=1:500-1000 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(chǎn)品介紹 Eukaryotic protein phosphorylation and dephosphorylation on serine and threonine residues regulates numerous cell functions, including division, homeostasis and apoptosis. A group of proteins that play a major role in this process are the serine/threonine protein phosphatases. Protein phosphatase (PP) holoenzyme is a trimeric complex that contains a regulatory subunit, a variable subunit and a catalytic subunit. Families of PP catalytic subunits include PP1, PP2A, PP2B, PP2C, PPX and PP5. Regulatory subunits include nuclear inhibitor of PP1 (NIPP1), PP1 nuclear targeting subunit (PNUTS), PP2A-A, PP2A-B, PP2A-B56, PP2A-C, PP2B-B and PR48. PNUTS, also designated CAT53 or FB19, is encoded by the gene PPP1R10. PNUTS acts as an inhibitor for the phosphatase activity of PP1 Alpha and PP1 Gamma. It is a nuclear protein primarily detected in nucleoplasmic bodies and within nucleoli. PNUTS expression levels are highest in brain, heart, lung, placenta, liver, kidney, pancreas and skeletal muscle.
Function : PPP1R10 (serine/threonine protein phosphatase 1 regulatory subunit 10), also known as PNUTS, is a targeting subunit of protein phosphatase 1 (PP1). Furthermore, PPP1R10 acts as an inhibitor of PPP1CA and PPP1CC phosphatase activities. Its inhibitory activity on PPP1CA is only observed when PPP1R10 is phosphorylated by Protein Kinase A (PKA). PPP1R10 contains a zinc finger motif and is predominantly localised within the nucleus. Recently, PPP1R10 has been shown to target PP1 to GABA(C) receptors, shuttling from the nucleus to the cell membrane in cells co-expressing this receptor. This highlights a role for this protein in the regulation of synaptic signal transduction. PPP1R10 has also been shown to regulate the activity of p53 in response to hypoxia.
Subunit : Component of the PTW/PP1 phosphatase complex, composed of PPP1R10/PNUTS, TOX4, WDR82, and PPP1CA or PPP1CB or PPP1CC. Interacts with PPP1CC. Interacts with PPP1CA, WDR82 and TOX4
Subcellular Location : Nucleus. Note=Found in discrete nucleoplasmic bodies and within nucleoli. Associates with chromatin during interphase, excluded from condensed chromosomes during early mitosis and is reloaded onto chromosomes at the late telophase
Post-translational modifications : Phosphorylated on Ser-398 and Thr-400 by PKA within the region necessary for interaction with PPP1CA
Similarity : Contains 1 C3H1-type zinc finger.
Contains 1 TFIIS N-terminal domain.
Database links : UniProtKB/Swiss-Prot: Q96QC0.1
抗體的鑒定:
1)絲氨酸/蘇氨酸蛋白磷酸酶1調(diào)節(jié)亞基10抗體說明書 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(jié)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(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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