SDS-PAGE: Coomassie-stained SDS-PAGE showing purified alpha-5 beta-3 protein.

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HUMAN INTEGRIN ALPHA-5 BETA-3, HIS-TAG
Recombinant human integrin alpha-5 beta-3 (ITGA5:ITGB3) protein produced in HEK293 cells and purified from culture supernatant.
PRODUCT DETAILS – HUMAN INTEGRIN ALPHA-5 BETA-3, HIS-TAG
- Recombinant human integrin ß-3 protein (NCBI accession number AAA52589.1, AA1-718) and human integrin α-5 (NCBI accession number NP_002196.4, AA1-992).
- Both proteins contain a C-terminal 6x histidine tag.
- Proteins were co-expressed in HEK293 cells and purified from culture supernatant by immobilised metal affinity chromatography.
- Resulting protein is a mix of integrin α-5 and integrin α-5/ß-3 heterodimer, sterile filtered in Dulbecco’s phosphate buffered saline pH7.4.
BACKGROUND
Integrins are heterodimeric integral membrane protein receptors composed of an alpha subunit and a beta subunit that function in cell surface adhesion and signaling. Each subunit crosses the membrane once, with most of the polypeptide residing in the extracellular space, and has two short cytoplasmic domains. A given chain may combine with multiple partners resulting in different integrins. Integrin alpha-5 or CD49e is a protein that in humans is encoded by the ITGA5 gene. Alpha chain 5 undergoes post-translational cleavage in the extracellular domain to yield disulfide-linked light and heavy chains that join with beta 1 to form a fibronectin receptor, recognizing the sequence R-G-D. The integrin alpha 5 and integrin alpha V subunits are encoded by distinct genes. Integrin alpha-V/beta-3 (ITGAV:ITGB3) is a receptor for cytotactin, fibronectin, laminin, matrix metalloproteinase-2, osteopontin, osteomodulin, prothrombin, thrombospondin, vitronectin and von Willebrand factor.
Numerous microorganisms utilize integrins to gain entry into cells. Integrin beta-3 (β3) or CD61 is a protein that in humans is encoded by the ITGB3 gene. Alpha V beta 3 is one of the most promiscuous integrins, as it binds to multiple ligands and also serves as a receptor for several viruses such as foot-and-mouth disease virus, adenovirus, and human immunodeficiency virus (Xiong et al., 2001).
Membrane fusion and attachment of pneumoviruses is achieved by the F protein which contains a putative integrin-binding motif (RGD) and binding to 5β1 and αv integrins are essential for cell-cell fusion and human metapneumovirus (hMPV) infection (Wei et al., 2014).
The specific receptors used for viruses within the Bunyaviridae remain mostly unknown; however, β3 and β1 integrins have been shown to be used as receptors for hantaviruses (Connolly-Andersen et al., 2007). It has been shown that β3 integrins facilitate the cellular entry of pathogenic hantavirus pulmonary syndrome (HPS) associated hantaviruses. Hantaviruses are enveloped viruses and their unique grid-like surface structure may require the use of integrins for entry into cells (Gavrilovskaya et al., 1998).
Other integrins have also been associated with virus entry into cells including ITGA5:ITGB1 which acts as a receptor for human metapneumovirus and as a coreceptor for parvovirus B19 (Weigel-Kelley et al., 2003). Integrin ITGAV:ITGB3 acts as a receptor for Herpes virus 8/HHV-8 (Garrigues et al., 2008), for Coxsackievirus A9 (Roivainen et al., 1994) and Cytomegalovirus/HHV-5 (Wang et al., 2005). Integrin ITGAV:ITGB3 also acts as a receptor for West Nile virus. Integrins were shown to modulate the infection efficiency of West Nile virus into cells and infection was substantially inhibited in Vero cells pretreated with blocking antibodies against αvβ3 integrin and its subunits by receptor competition assay. However, the binding of WNV to αvβ3 integrin does not specifically occur on the RGD binding site. Cells pretreated with antibodies against αvβ3 integrin also inhibited flavivirus Japanese encephalitis and to a lesser extent flavivirus dengue infections (Chu and Ng, 2004; Schmidt et al., 2013).
Antibodies against the receptors αvβ3 and αvβ5 block adenovirus internalization without affecting attachment. Additionally, adenovirus binds to cultured cells lacking αv integrins but fail to become internalized (Wickham et al., 1993).
Integrins may also determine the pathway of virus entry. Herpes Simplex virus (HSV) enters CHO cells lackingαVβ3-integrin through a pathway independent of cholesterol-rich rafts and dynamin2. In the presence of αVβ3-integrin, HSV enters CHO-nectin1 cells through a pathway dependent on cholesterol-rich rafts and dynamin2 (Gianni et al., 2010).
REFERENCES
- Chu and Ng (2004). Interaction of West Nile virus with alpha v beta 3 integrin mediates virus entry into cells. J Biol Chem. 279(52):54533-41.
- Connolly-Andersen et al. (2007). Basolateral entry and release of Crimean-Congo hemorrhagic fever virus in polarized MDCK-1 cells. J Virol. 81(5):2158-64.
- Garrigues et al. (2008). Integrin alphaVbeta3 Binds to the RGD motif of glycoprotein B of Kaposi’s sarcoma-associated herpesvirus and functions as an RGD-dependent entry receptor. J Virol. 82(3):1570-80.
- Gavrilovskaya et al. (1998). beta3 Integrins mediate the cellular entry of hantaviruses that cause respiratory failure. Proc Natl Acad Sci U S A. 95(12):7074-9.
- Gianni et al. (2010). αVβ3-integrin routes herpes simplex virus to an entry pathway dependent on cholesterol-rich lipid rafts and dynamin2. PNAS 107 (51) 22260-22265.
- Horton MA (1997). The alpha v beta 3 integrin vitronectin receptor. Int J Biochem Cell Biol. 29(5):721-5.
- Roivainen et al. (1994). Entry of coxsackievirus A9 into host cells: specific interactions with alpha v beta 3 integrin, the vitronectin receptor. Virology. 1994 Sep;203(2):357-65.
- Schmidt et al. (2013). Integrin ITGAV:ITGB3 acts as a receptor for West nile virus. J Gen Virol. 94(Pt 8):1723-33.
- Wang et al. (2005). Integrin alphavbeta3 is a coreceptor for human cytomegalovirus. Nat Med. 11(5):515-21.
- Wei et al. (2014). Roles of the putative integrin-binding motif of the human metapneumovirus fusion (f) protein in cell-cell fusion, viral infectivity, and pathogenesis. J Virol. 88(8):4338-52.
- Weigel-Kelley et al. (2003). Alpha5beta1 integrin as a cellular coreceptor for human parvovirus B19: requirement of functional activation of beta1 integrin for viral entry. Blood. 102(12):3927-33.
- Wickham et al. (1993). Integrins alpha v beta 3 and alpha v beta 5 promote adenovirus internalization but not virus attachment. Cell. 73(2):309-19.
- Xiong et al. (2001). Crystal Structure of the Extracellular Segment of Integrin αVβ3. Science. 294(5541): 339–345.
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①乙型肝炎表面抗原—抗体系统(HBsAg/抗—HBs);
②乙型肝炎核心抗原—抗体系统(HBcAg/抗—HBc):
③乙型肝炎e抗原—抗体系统(HBeAg);
④乙型肝炎Dane颗粒抗原—抗体系统
⑤乙型肝炎δ抗原—抗体系统(δ/抗—δ)。
临床意义 1.HBsAg:血清中检测到HBsAg ,表示体内感染了HBV,因而是一种特异性标志。HBsAg阳性见于:①急性乙型肝炎的潜伏期或急性期(大多短期阳性);②HBV致的慢性肝病、迁延性和慢性活动性肝炎、肝炎后肝硬化或原发性肝癌等。③无症状携带者
2.抗HBs:表示曾感染过HBV,不论临床上有无肝炎症状表现,均已得到恢复,并且对HBV有一定的免疫力。
3.HBcAg与抗HBc:由于 HBcAg主要存在于肝细胞核内,并仅存在于Dane颗粒中。因此,对病人血清不能检测HBcAg,而测抗HBc。血清内抗HBc阳性反映:①新近有过HBV感染;②体内有HBV增殖;③有助于诊断急性或慢性乙型肝炎,特别是少数病例就诊时已处于急性恢复期早期,HBsAg已从血中消失,此时血中仅有抗HBc存在,因此,对恢复期患者可作病因追索。
4.HBcAg和抗HBe:HBcAg的存在常表示病人血液有感染性。 HBcAg阳性揭示病人肝脏可能有慢性损害,对预后判断有一定帮助。抗HBe阳性对病人可能有一定的保护力。展开
流感病毒的基因组包含________基因,其中__________蛋白可作为__________的结构基础.
我不知道第一空如何填,第二空是否应该填"HA蛋白"?第3空是否应该填"突变"?
多谢各位啊!
新年快乐!
万事如意!
主要是中度以上细胞免疫缺陷包括:CD4+T淋巴细胞耗竭,外周血淋巴细胞显著减少,CD4<200/μl,CD4/CD8<1.0,(正常人为1.25~2.1),迟发型变态反应皮试阴性,有丝分裂原刺激反应低下。NK细胞活性下降。
2.各种致病性感染的病原体检查
如用PCR方法检测相关病原体,恶性肿瘤的组织病理学检查。
3.HIV抗体检测
采用酶联免疫吸附法、明胶颗粒凝集试验、免疫荧光检测法、免疫印迹检测法、放射免疫沉淀法等,其中前三项常用于筛选试验,后二者用于确证试验。
4.PCR技术检测HIV病毒。
核心抗原由183个或185个氨基酸组成,高度磷酸化,是乙肝病毒核心颗粒的唯一结构蛋白。正由于它存在于Dane颗粒核心结构表面,被表面抗原覆盖,故不易在血循环中检出。核心抗原具有强免疫原性,可诱导很强的体液免疫和细胞免疫,刺激机体产生抗-HBc。
e抗原为可溶性蛋白质,传染性强,游离存在于血液中,虽然很早就被发现,在病理上认为是HBV复制以具有强感染性的一个指标,但其功能尚不清楚。抗-HBe的出现,是预后良好的征象。向左转|向右转
1、存在于细胞外环境时,不显复制活性,但保持感染活性,是病毒体或病毒颗粒形式。
2、抗原,是指能够刺激机体产生(特异性)免疫应答,并能与免疫应答产物抗体和致敏淋巴细胞在体外结合,发生免疫效应(特异性反应)的物质