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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 | CS9795 |
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 ICC=1:100-500 IF=1:50-200 |
CAS編號 | |
抗體名 | Anti-phospho-NIPA(Ser354) |
克隆性 | |
靶點 | 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 |
標識物 | 詳見說明書 |
濃度 | 1mg/1ml% |
免疫原 | KLH conjugated synthesised phosphopeptide derived from human NIPA around the phosphorylation site of Ser354 |
抗體的生物素化標記實驗要點:
1. 磷酸化間變性激酶核相互作用伴侶蛋白抗體品牌 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,選擇不當則影響標記的效率,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*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-phospho-NIPA(Ser354)
中文名稱 磷酸化間變性激酶核相互作用伴侶蛋白抗體品牌
別 名 NIPA (phospho S354); NIPA (phospho Ser354); p-NIPA (S354); p-NIPA (Ser354); hNIPA; Nuclear interacting partner of ALK; Nuclear interacting partner of anaplastic lymphoma kinase; ZC3HC1; Zinc finger C3HC type containing 1; NIPA_HUMAN.
濃 度 1mg/1ml
規(guī) 格 0.1ml/100μg
抗體來源 Rabbit
克隆類型 polyclonal
交叉反應(yīng) Human, Mouse, Rat, Dog, Pig, Horse, Rabbit
產(chǎn)品類型 一抗 磷酸化抗體
研究領(lǐng)域 細胞生物 免疫學 染色質(zhì)和核信號 信號轉(zhuǎn)導 激酶和磷酸酶
蛋白分子量 predicted molecular weight: 55kDa
性 狀 Lyophilized or Liquid
免 疫 原 KLH conjugated synthesised phosphopeptide derived from human NIPA around the phosphorylation site of Ser354
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
磷酸化間變性激酶核相互作用伴侶蛋白抗體品牌 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 ICC=1:100-500 IF=1:50-200
(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 regulated oscillation of protein expression is an essential mechanism of cell cycle control. The SCF class of E3 ubiquitin ligases is involved in this process by targeting cell cycle regulatory proteins for degradation by the proteasome, with the F-box subunit of the SCF specifically recruiting a given substrate to the SCF core. NIPA (nuclear interaction partner of ALK) is a human F-box-containing protein that defines an SCF-type E3 ligase (SCFNIPA) controlling mitotic entry. Assembly of this SCF complex is regulated by cell-cycle-dependent phosphorylation of NIPA, which restricts substrate ubiquitination activity to interphase. Nuclear cyclin B1 is a substrate of SCFNIPA. Inactivation of NIPA by RNAi results in nuclear accumulation of cyclin B1 in interphase, activation of cyclin B1-Cdk1 kinase activity, and premature mitotic entry. Thus, SCFNIPA-based ubiquitination may regulate S-phase completion and mitotic entry in the mammalian cell cycle.
Function : Essential component of an SCF-type E3 ligase complex, SCF(NIPA), a complex that controls mitotic entry by mediating ubiquitination and subsequent degradation of cyclin B1 (CCNB1). Its cell-cycle-dependent phosphorylation regulates the assembly of the SCF(NIPA) complex, restricting CCNB1 ubiquitination activity to interphase. Its inactivation results in nuclear accumulation of CCNB1 in interphase and premature mitotic entry. May have an antiapoptotic role in NPM-ALK-mediated signaling events.Subunit : Interacts with the NPM-ALK fusion protein in a tyrosine phosphorylation-dependent manner. Interacts with SKP1. Component of a SCF(NIPA) E3 complex with SKP1, RBX1 and CUL1 when not phosphorylated on Ser-354. Interacts with CCNB1.Subcellular Location : Nuclear.Tissue Specificity : Widely expressed. Highly expressed in heart, skeletal muscle and testis. Expressed in brain, placenta, lung, kidney, liver, pancreas, spleen, thymus, prostate, ovary small intestine and colon. Weakly or not expressed in leukocytes.Post-translational modifications : hosphorylated. Phosphorylated on Ser residues at G2/M phase, but not during S and G0 phases. May also be weakly phosphorylated on Tyr residues. Ser-354 phosphorylation, a major site during the course of cell-cycle-dedendent phosphorylation, results in its dissociation from the SCF(NIPA) complex, thereby preventing CCNB1 degradation leading to mitotic entry.Similarity : Contains 1 C3HC-type zinc finger.Database links : UniProtKB/Swiss-Prot: Q86WB0.1
抗體的鑒定:
1)磷酸化間變性激酶核相互作用伴侶蛋白抗體品牌 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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IgG 干粉 10mg/50mg/100mg 國產(chǎn)
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Delta 4 英文名稱: Notch信號通路Delta樣配體4抗體 0.1ml
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MGLUR2 + MGLUR3 英文名稱: 促代謝型谷酸受體2+3抗體 0.2ml
ADCK2 英文名稱: ADCK2蛋白抗體 0.2ml
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AQP-2 (aquaporin Protein-2) 水通道蛋白-2(抗原) 0.5mg
GABA A Receptor alpha 1 英文名稱: G1基A型受體α1抗體 0.1ml
Rhesus antibody Rh Rabbit Anti-rat IgM/Cy7 Cy7標記的兔抗大鼠IgM 規(guī)格 0.1ml
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Bordet-GengouAgarMedium
DMEM(低糖),液體 "含1000mg/l葡萄糖,L-谷氨酰胺, incubation media DMEM(低糖),液體 "含1000mg/l葡萄糖,L-谷氨酰胺,
膠質(zhì)芽孢桿菌 支/瓶
NA平板(加青霉素酶)(9cm)細菌計數(shù)、不含糖,可作血瓊脂基礎(chǔ)和傳代用
XLDAgar
ThioglycollateMedium
馬鈴薯-葡萄糖瓊脂培養(yǎng)基(不含氯霉素) 250(g) incubation media 馬鈴薯-葡萄糖瓊脂培養(yǎng)基(不含氯霉素) 250(g)
察氏培養(yǎng)基 Czapek Dox Agar 250克 用于酵母菌的培養(yǎng)
多粘菌素B2.5萬單位/支*5添加于HB4incubationmedia多粘菌素B2.5萬單位/支*5添加于HB4131中
BufferedPeptoneWater
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