- SynonymILT1,LIR7,CD85H,LIR-7,LILRA2,ILT1CD85H,CD85h,ILT1,ILT-1,ILT1CD85H,CD85h antigen,CD85 antigen-like family member H
- SourceBiotinylated Human LILRA2, His,Avitag (LI2-H82E9) is expressed from human 293 cells (HEK293). It contains AA Gly 24 - Asn 449 (Accession # Q8N149-1).Predicted N-terminus: Gly 24Request for sequence
- Molecular Characterization
This protein carries a polyhistidine tag at the C-terminus, followed by an Avi tag.
The protein has a calculated MW of 50.7 kDa. The protein migrates as 67-100 kDa under reducing (R) condition (SDS-PAGE) due to glycosylation.
- BiotinylationBiotinylation of this product is performed using Avitag™ technology. Briefly, the single lysine residue in the Avitag is enzymatically labeled with biotin.
- EndotoxinLess than 1.0 EU per μg by the LAL method.
- Purity
>90% as determined by SDS-PAGE.
- Formulation
Lyophilized from 0.22 μm filtered solution in PBS, pH7.4. Normally trehalose is added as protectant before lyophilization.
Contact us for customized product form or formulation.
- Reconstitution
Please see Certificate of Analysis for specific instructions.
For best performance, we strongly recommend you to follow the reconstitution protocol provided in the CoA.
- Storage
For long term storage, the product should be stored at lyophilized state at -20°C or lower.
Please avoid repeated freeze-thaw cycles.
This product is stable after storage at:
- -20°C to -70°C for 12 months in lyophilized state;
- -70°C for 3 months under sterile conditions after reconstitution.
Biotinylated Human LILRA2, His,Avitag on SDS-PAGE under reducing (R) condition. The gel was stained overnight with Coomassie Blue. The purity of the protein is greater than 90%.
Immobilized Human ANGPTL7, His Tag (Cat. No. AN7-H52H5) at 5 μg/mL (100 μL/well) can bind Biotinylated Human LILRA2, His,Avitag (Cat. No. LI2-H82E9) with a linear range of 0.625-5 μg/mL (QC tested).
- BackgroundLILRA2 (Leukocyte immunoglobulin like receptor A2), also known as CD85H and LIR7 (Leukocyte immunoglobulin-like receptor 7). It belongs to the subfamily A class of LIR receptors (LILRAs), which are single-pass type I transmembrane proteins with 2-4 extracellular Ig-like domains, a transmembrane domain (TM), and a short cytoplasmic tail.LILRA2 contains 4 Ig-like C2 type domains in the extracellular region. LILRA2 does not bind class I MHC antigens. LILRA2 is expressed predominantly on monocytes and B cells, and at lower levels on dendritic cells and natural killer cells. LILRA2 is an activating receptor that inhibits dendritic cell differentiation and antigen presentation and suppresses innate immune response.
- References
- (1)Hirayasu K., et al., 2016, Nat Microbiol.,1(6), 16054.
- (2)Brown D., et al., 2004, Tissue Antigens.,64(3), 215-25.
- (3)MacGlashan DW Jr1., et al.,2008, J Immunol., 180(6), 4208-17.
Please contact us via TechSupport@acrobiosystems.com if you have any question on this product.
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(一)包被用抗原
用于包被固相载体的抗原按其来源不同可分为天然抗原、重组抗原和合成多肽抗原三大类。天然抗原可取自动物组织、微生物培养物等,须经提取纯化才能作包被用。如HBsAg可以从携带者的血清中提取,一般的细菌和病毒抗原可以从其培养物中提取,蛋白成份抗原可从富含此抗原的材料中提取等(例如AFP从脐带血或胎肝中提取)。重组抗原是抗原基因在质粒体中表达的蛋白质抗原,多以大肠杆菌或酵母菌为质粒体。重组抗原的优点是除工程菌成份外,其他杂质少,而且无传染性,但纯化技术难度较大。以大肠杆菌为质粒体的重组抗原如不能充分除大肠杆菌成份,用于ELISA试剂盒,在反应中可出现假阳性,因不少受检者受大肠杆菌感染而在血清中存在抗大肠杆菌抗体。重组抗原的另一特点是能用基因工程制备某些无法从天然材料中分离的抗原物质。例如丙型肝炎病毒(HCV)尚不能培养成功,而且丙肝病人血清中HCV抗原含量极微。目前检测抗HCVELISA中所用包被抗原大多为根据HCV的基因克隆表达而制备的重组抗原。在传染病诊断中,不少重组抗原如HBsAg、HBeAg和HIV抗原等均在ELISA中取得应用。合成多肽抗原是根据蛋白质抗原分子的某一抗原决定簇的氨基酸序列人工合成的多肽片段。多肽抗原一般只含有一个抗原决定簇,纯度高,特异性也高,但由于分子量太小,往往难于直接吸附于固相上。多肽抗原的包被一般需先使其与无关蛋白质如牛血清白蛋白质(BSA)等偶联,借助于偶联物与固相载体的吸附,间接地结合到固相载体表面。应用多肽抗原的另一注意点为他仅能检测与其相应的抗体。一种蛋白质抗原往往含有多个不同的能引起抗体产生的决定簇,因此在受检血清中的其他抗体就不能与该多肽抗原发生反应。另外,某些微生物发生变异时往往发生抗原结构变化,在这种情况下,用个别多肽抗原进行包被可引起其他抗体的漏检。
(二)ELISA试剂盒包被用抗体
包被固相载体的抗体应具有高亲和力和高特异性,可取材于抗血清或含单克隆抗体的腹水或培养液。如免疫用抗原中含有杂质(即便是极微量的),在抗血清中将出现杂抗体,必须除去(可用吸收法)后才能用于ELISA,以保证试验的特异性。抗血清不能直接用于包被,应先提取IgG,通常采用硫酸铵盐析和Sephadex凝胶过滤法。一般经硫酸铵盐析粗提的IgG已可用于包被,高度纯化的IgG性质不稳定。如需用高亲和力的抗体包被以提高试验的敏感性,则可采用亲和层析法以除去抗血清中含量较多的非特异性IgG。腹水中单抗的浓度较高,特异性亦较强,因此不需要作吸收和亲和层析处理,一般可将腹水作适当稀释后直接包被,必要时也可用纯化的IgG。应用单抗包被时应注意,一种单抗仅针对一种抗原决定簇,在某些情况下,用多种单抗混合包被,可取得更好的效果。展开
补充一点的是有些病毒在人群中的感染是很普遍的,比如HCMV.对于这些病毒IgM的检测结果可以推测是否为近期感染.但这一类病毒其检测的临床意义经常是当其再激活感染时.比如:HCMV感染后就在末梢单个核细胞中潜伏起来,对于正常人而言是不致病的,但当出现HIV感染或应用免疫抑制剂时会出现激活感染而且经常导致严重疾患.对于这种再激活感染虽然也有IgM的升高,但是却在现在ELISA技术的检测限以下,因此是检测不到的,这时就必须进行针对其抗原的检测。
双抗无论是在免疫学还是细胞操作,细胞培养中都不与二抗等同。双抗可以是抗一种抗原的两个抗体,去双抗夹心法。也可以是抗不同类型反应的作用物,如抗凝剂素。一般指两种抗生素。
液体的一抗加50%甘油,-20度也不会冻结,可保存3-5年;
经常使用时取出一小份,4度可使用半年之久;
粉末状的二抗直接-20或-80度贮藏就行了。
一抗的英语是 resistance No.1,二抗resistance No.2.
我的实验是这样的,先纯化质粒,然后做酶切,再纯化DNA。
最后一步其实也可以用切胶回收试剂盒,我怕做不好,所以想选用DNA纯化试剂盒,但不知跟质粒DNA提取试剂盒是否有区别,因为说明书上说可以用Invitrogen的K210001.我搜索看了一席啊,觉得那个是提取质粒的小提啊。所以产生这个疑问,请大侠告知DNA纯化试剂盒与质粒DNA提取试剂盒这两者之间有区别吗?
用哪个公司的DNA纯化试剂盒比较好。谢谢啦!
想通过亲和层析来纯化抗血小板特异性抗体(抗GPIIb/IIIa抗体和抗GPIbα抗体),能否做到呢?谢谢!
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