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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 | CS11878 |
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-Phospho-Tuberin/TSC2 (Ser1254) |
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(biāo)識物 | 詳見說明書 |
濃度 | 1mg/1ml% |
免疫原 | KLH conjugated Synthesised phosphopeptide derived from human Tuberin around the phosphorylation site of Ser1254 [SL(p-S)VP] |
抗體的生物素化標(biāo)記實驗要點:
1. 磷酸化馬鈴薯球蛋白(性硬化)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(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-氨基的密度會有所不同,選擇不當(dāng)則影響標(biāo)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(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(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(xì)胞的熒光標(biāo)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(xì)胞可導(dǎo)致高本底。
產(chǎn)品訂購信息:
英文名稱 Anti-Phospho-Tuberin/TSC2 (Ser1254)
中文名稱 磷酸化馬鈴薯球蛋白(性硬化)抗體規(guī)格
別 名 FLJ43106; LAM; TSC2; TSC2_HUMAN; TSC4; Tuberin; Tuberous sclerosis 2 protein.
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
產(chǎn)品類型 一抗 磷酸化抗體
研究領(lǐng)域 心 細(xì)胞生物 免疫學(xué) 神經(jīng)生物學(xué) 細(xì)胞凋亡
蛋白分子量 predicted molecular weight: 200kDa
性 狀 Lyophilized or Liquid
免 疫 原 KLH conjugated Synthesised phosphopeptide derived from human Tuberin around the phosphorylation site of Ser1254 [SL(p-S)VP]
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
磷酸化馬鈴薯球蛋白(性硬化)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(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 Tuberin, or TSC2 (Tuberous sclerosis complex), is implicated as a tumor suppressor. It may function in vesicular transport, and may also play a role in the regulation of cell growth arrest and in the regulation of transcription mediated by steroid receptors. Interaction between hamartin (TSC1) and tuberin may facilitate vesicular docking. It specifically stimulates the intrinsic GTPase activity of the Ras related protein RAP1A and RAB5, suggesting a possible mechanism for its role in regulating cellular growth. Mutations in tuberin lead to constitutive activation of RAP1A in tumors. At least three isoforms of Tuberin exist.
Function : In complex with TSC1, inhibits the nutrient-mediated or growth factor-stimulated phosphorylation of S6K1 and EIF4EBP1 by negatively regulating mTORC1 signaling. Acts as a GTPase-activating protein (GAP) for the small GTPase RHEB, a direct activator of the protein kinase activity of mTORC1. Implicated as a tumor suppressor. Involved in microtubule-mediated protein transport, but this seems to be due to unregulated mTOR signaling. Stimulates weakly the intrinsic GTPase activity of the Ras-related proteins RAP1A and RAB5 in vitro. Mutations in TSC2 lead to constitutive activation of RAP1A in tumors.
Subunit : Interacts with TSC1 and HERC1; the interaction with TSC1 stabilizes TSC2 and prevents the interaction with HERC1. May also interact with the adapter molecule RABEP1. The final complex contains TSC2 and RABEP1 linked to RAB5 (Probable). Interacts with HSPA1 and HSPA8. Interacts with DAPK1.
Subcellular Location : Cytoplasm. Membrane; Peripheral membrane protein. Note=At steady state found in association with membranes.
Tissue Specificity : Liver, brain, heart, lymphocytes, fibroblasts, biliary epithelium, pancreas, skeletal muscle, kidney, lung and placenta.
Post-translational modifications : Phosphorylation at Ser-1387, Ser-1418 or Ser-1420 does not affect interaction with TSC1. Phosphorylation at Ser-939 and Thr-1462 by PKB/AKT1 is induced by growth factor stimulation. Phosphorylation by AMPK activates it and leads to negatively regulates the mTORC1 complex. Phosphorylated at Ser-1798 by RPS6KA1; phosphorylation inhibits TSC2 ability to suppress mTORC1 signaling. Phosphorylated by DAPK1.
DISEASE : Defects in TSC2 are the cause of tuberous sclerosis type 2 (TSC2) [MIM:613254]. TSC2 is an autosomal dominant multi-system disorder that affects especially the brain, kidneys, heart, and skin. It is characterized by hamartomas (benign overgrowths predominantly of a cell or tissue type that occurs normally in the organ) and hamartias (developmental abnormalities of tissue combination). Clinical symptoms can range from benign hypopigmented macules of the skin to profound mental retardation with intractable seizures to premature death from a variety of disease-associated causes.
Defects in TSC2 are a cause of lymphangioleiomyomatosis (LAM) [MIM:606690]. LAM is a progressive and often fatal lung disease characterized by a diffuse proliferation of abnormal smooth muscle cells in the lungs. It affects almost exclusively young women and can occur as an isolated disorder or in association with tuberous sclerosis complex.
Database links : UniProtKB/Swiss-Prot: P49815.2
Tuberin 為180kD-220的蛋白產(chǎn)生于性硬化(TSC-2)位于染色體16的基因。性硬化為常染色體疾病已知為斑痣性錯構(gòu)瘤病其特征為廣泛性發(fā)展呈良性生長,在許多組織與器官中描述為錯構(gòu)瘤。Tuberin被認(rèn)為在GTPase活動性蛋白中起作用調(diào)節(jié)細(xì)胞內(nèi)吞與作為抑制子。Tuberin低水平廣泛性表達在大多數(shù)組織中,而在皮層神經(jīng)元、小腦浦肯野氏細(xì)胞、脊索運動神經(jīng)元、胰島B細(xì)胞、心肌、*與皮膚的小中表達增加。
抗體的鑒定:
1)磷酸化馬鈴薯球蛋白(性硬化)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(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。抗體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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