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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 | CS9846 |
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-Nanog |
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 synthetic peptide derived from mouse Nanog |
抗體的生物素化標記實驗要點:
1. 胚胎干細胞關(guān)鍵蛋白抗體規(guī)格 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-Nanog
中文名稱 胚胎干細胞關(guān)鍵蛋白抗體規(guī)格
別 名 NANOG_MOUSE; Embryonic stem cell specific homeobox protein (Nanog); ENK; FLJ12581; FLJ40451; Homeobox transcription factor Nanog; Nanog homeobox; Homeobox protein NANOG; ES cell-associated protein 4; Early embryo specific expression NK-type homeobox protein; Homeobox transcription factor Nanog; Nanog; Ecat4.
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) Mouse, Rat, Pig
產(chǎn)品類型 一抗
研究領(lǐng)域 干細胞
蛋白分子量 predicted molecular weight: 34kDa
性 狀 Lyophilized or Liquid
免 疫 原 KLH conjugated synthetic peptide derived from mouse Nanog
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
胚胎干細胞關(guān)鍵蛋白抗體規(guī)格 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)
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 Nanog is a newly identified homeodomain-bearing transcriptional factor. Nanog expression is specific to early embryos and pluripotential stem cells including mouse and human embryonic stem (ES) and embryonic germ (EG) cells. It is a key molecule involved in the signaling pathway for maintaining the capacity for self-renewal and pluripotency, bypassing regulation by the STAT3 pathway. Nanog mRNA is present in pluripotent mouse and human cell lines, and absent from differentiated cells. Nanog-deficient ES cells lose pluripotency and differentiate into extraembryonic endoderm lineage. Thus it is one of the molecular markers suitable for recognizing the undifferentiated state of stem cells in the mouse and human.
NANOG is a new marker for testicular carcinoma in situ and germ cell tumors.NANOG is a gene expressed in embryonic stem cells (ESCs) and is thought to be a key factor in maintaining pluripotency. NANOG thought to function in concert with other factors such as POU5F1 and SOX2 to establish ESC identity. These cells offer an important area of study because of their ability to maintain pluripotency. In other words, these cells have the ability to become virtually any cell of any of the three germ layers (endoderm, ectoderm, mesoderm).Function : Transcription regulator involved in inner cell mass and embryonic stem (ES) cells proliferation and self-renewal. Imposes pluripotency on ES cells and prevents their differentiation towards extraembryonic endoderm and trophectoderm lineages. Blocks bone morphogenetic protein-induced mesoderm differentiation of ES cells by physically interacting with SMAD1 and interfering with the recruitment of coactivators to the active SMAD transcriptional complexes. Acts as a transcriptional activator or repressor. Binds optimally to the DNA consensus sequence 5'-TAAT[GT][GT]-3' or 5'-[CG][GA][CG]C[GC]ATTAN[GC]-3'. When overexpressed, promotes cells to enter into S phase and proliferation.Subunit : Interacts with SMAD1 and SALL4.Subcellular Location : Nucleus.Tissue Specificity : Expressed in testicular carcinoma and derived germ cell tumors (at protein level). Expressed in fetal gonads, ovary and testis. Also expressed in ovary teratocarcinoma cell line and testicular embryonic carcinoma. Not expressed in many somatic organs and oocytes.Similarity : Belongs to the Nanog homeobox family.Contains 1 homeobox DNA-binding domain.Database links :UniProtKB/Swiss-Prot: Q80Z64.1Entrez Gene: 100293888 HumanEntrez Gene: 79923 HumanOmim: 607937 HumanSwissProt: Q9H9S0 HumanUnigene: 635882 Human研究表明;早期胚胎在沒有Nanog存在的情況下是不能存活的,但是這一蛋白的表達水平至今還有待于進一步研究。學者認為:Nanog依然是一種對于生殖細胞(germ lls)形成的必要因子,Nanog也是調(diào)控著細胞融合后的多重功能的關(guān)鍵因子。胚胎干細胞是當今生命科學和生物技術(shù)研究的熱點,這主要是由于其具有“發(fā)育全能性”的功能。ES能分化成人體200多種細胞類型,形成機體的任何細胞、組織和器官。通過掌握其分化發(fā)育的規(guī)律,在人工條件下定向分化為所需的細胞、組織乃至器官,科學家們希望可以用來目前還難以或無法的、、、等頑癥,并且解決十分緊缺的組織和的來源問題,并且通過進一步與克隆技術(shù)相結(jié)合,運用體細胞核轉(zhuǎn)移技術(shù)來得到ES,還能解決細胞以及組織和的免疫排異難題。
抗體的鑒定:
1)胚胎干細胞關(guān)鍵蛋白抗體規(guī)格 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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