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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 | CS9833 |
應用范圍 | WB=1:100-500 ELISA=1:500-1000 IHC-P=1:100-500 IHC-F=1:100-500 Flow-Cyt=1:100-500 ICC=1:100-500 IF=1:100-500 |
CAS編號 | |
抗體名 | Anti-NG2/CSPG4 |
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 NG2 |
抗體的生物素化標記實驗要點:
1. 硫酸軟骨素蛋白多糖4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*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-NG2/CSPG4
中文名稱 硫酸軟骨素蛋白多糖4抗體費用
別 名 MELCSPG; AN2; AN2 proteoglycan; Chondroitin sulfate proteoglycan 4 (melanoma-associated); Chondroitin sulfate proteoglycan 4; Chondroitin sulfate proteoglycan NG2; CSPG4; Cspg4 chondroitin sulfate proteoglycan 4; CSPG4_HUMAN; HMW-MAA; HSN tumor-specific antigen; Kiaa4232; MCSP; MCSPG; MEL-CSPG; Melanoma chondroitin sulfate proteoglycan; Melanoma-associated chondroitin sulfate proteoglycan; 4732461B14Rik; MSK16.
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, Pig, Cow
產(chǎn)品類型 一抗
研究領域 細胞生物 免疫學 神經(jīng)生物學
蛋白分子量 predicted molecular weight: 247kDa
性 狀 Lyophilized or Liquid
免 疫 原 KLH conjugated synthetic peptide derived from human NG2
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
硫酸軟骨素蛋白多糖4抗體費用 產(chǎn)品應用 ELISA=1:500-1000 IHC-P=1:100-500 IHC-F=1:100-500 Flow-Cyt=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 NG2 (also known as melanoma-associated chondroitin sulfate proteoglycan 4, MCSP, MCSPG, MSK16 and MEL-CSPG) stabilizes cell-substratum interactions during early events of melanoma cell spreading on endothelial basement membranes. NG2 may facilitate primary melanoma progression by enhancing the activation of key signaling pathways important for tumor invasion and growth. Threonine 2256 phosphorylation of rat NG2 (Threonine 2252 phosphorylation of human NG2) leads to redistribution of NG2 on the surface of astrocytomas, polarization of the cell and a significant increase in cell motility. NG2 acts as a co-receptor for spreading and focal contact formation in association with Alpha 4 Beta1 integrin in malignant melanoma cells. NG2 is present on blood vessels throughout the rat embryo. Microvessels within the rat CNS express NG2 on endothelial cells, and outside the CNS, NG2 is present on smooth muscle cells. NG2 is a novel marker for epidermal stem cells that contributes to their patterned distribution by promoting stem cell clustering.
Function : Detected only in malignant melanoma cells.Subunit : Interacts with the first PDZ domain of MPDZ. Interacts with PRKCA. Binds TNC, laminin-1, COL5A1 and COL6A2. Interacts with PLG and angiostatin. Binds FGF2 and PDGFA. Interacts with GRIP1, GRIP2 and GRIA2. Forms a ternary complex with GRIP1 and GRIA2 (By similarity). Interacts with LGALS3 and the integrin composed of ITGB1 and ITGA3. Interacts with ITGA4 through its chondroitin sulfate glycosaminoglycan. Interacts with BCAR1, CDC42 and ACK1. Interacts with MMP16.Subcellular Location : Apical cell membrane. Cell projection > lamellipodium membrane. Localized at the apical plasma membrane it relocalizes to the lamellipodia of astrocytoma upon phosphorylation by PRKCA. Localizes to the retraction fibers. Localizes to the plasma membrane of oligodendrocytes.Tissue Specificity : Detected only in malignant melanoma cells.Post-translational modifications : O-glycosylated; contains glycosaminoglycan chondroitin sulfate which are required for proper localization and function in stress fiber formation (By similarity). Involved in interaction with MMP16 and ITGA4.Phosphorylation by PRKCA regulates its subcellular location and function in cell motility (By similarity).Similarity : Contains 15 CSPG (NG2) repeats.Contains 2 laminin G-like domains.Database links : UniProtKB/Swiss-Prot: Q6UVK1.2
抗體的鑒定:
1)硫酸軟骨素蛋白多糖4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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