Clone | MON161 |
Isotype | IgG1 |
Product Type | Primary Antibodies |
Units | 0.1 mg |
Host | Mouse |
Species reactivity | HamsterHumanMouseRat |
Application | Flow CytometryImmunoblottingImmunocytochemistryImmunohistochemistry (frozen)Immunohistochemistry (paraffin)Western Blotting |
BackgroundRecently, a novel gene family has been identified and characterized, designated the Reticulons because the proteins encoded by these genes are anchored to the membranes of the endoplasmic reticulum. Reticulon-1 was formerly designated NSP for Neuroendocrine-Specific-Protein, because it is specifically expressed in neural and neuroendocrine tissues. The NSP-gene has been mapped by fluorescence in situ hybridization to Human chromosome 14q21-q22. The NSP-gene encodes three overlapping proteins, i.e. Reticulon-1A (NSP-A), Reticulon-1B (NSP-B), and Reticulon-1C (NSP-C). These proteins were found to be anchored to membranes of the endoplasmic reticulum through their common carboxy-terminal regions. Reticulon-1A is a protein with a molecular weight (MW) of about 135 kDa, which occurs in various isoforms presumably depending on the degree of phosphorylation of serine residues. In lung cancer diagnosis Reticulon-1A appeared to be a reliable marker for the detection of neuroendocrine differentiation, since most of the small cell lung carcinoma (SCLC) and carcinoid tumors showed expression of Reticulon-1A. Reticulon-1B is a phosphoprotein with a MW of 45 kDa and is restricted to the lung cancer cell line NCI-H82. Reticulon-1B is sofar not found in Human tissues. Reticulon-1C is a protein with a MW of 23 kDa which is not phosphorylated and is found with Reticulon-1A in SCLC (cell lines) and not in non-SCLC (cell cultures).
SourceMON-161 is a Mouse monoclonal IgG1 antibody derived by fusion of Mouse myeloma cells with spleen cells from a Mouse immunized with a partially purified bacterially expressed Reticulon-1A (NSP-A) hybrid protein (βGAL-NSP-A 6-776).
ProductEach vial contains 100 ul 1 mg/ml purified monoclonal antibody in PBS containing 0.09% sodium azide.
Formulation: Each vial contains 100 ul 1 mg/ml purified monoclonal antibody in PBS containing 0.09% sodium azide.
SpecificityMON-161 exclusively recognizes the 135 kD Reticulon-1A protein in immunoblots of NCI-H82 and other SCLC cell lines, and stains normal and pathological neural and neuroendocrine tissues. The epitope of MON-161 is loCated between amino acid residues 174-337 of Reticulon-1A.
ApplicationsMON-161 is useful for immunocytochemistry, immunohistochemistry on frozen and paraffin-embedded tissue, immunoblotting and flow cytometry. Optimal antibody dilution should be determined by titration; recommended range is 1:50 – 1:100 for flow cytometry, and for immunohistochemistry with avidin-biotinylated Horseradish peroxidase complex (ABC) as detection reagent, and 1:50 – 1:500 for immunoblotting applications.
StorageThe antibody is shipped at ambient temperature and may be stored at +4°C. For prolonged storage prepare appropriate aliquots and store at or below -20°C. Prior to use, an aliquot is thawed slowly in the dark at ambient temperature, spun down again and used to prepare working dilutions by adding sterile phosphate buffered saline (PBS, pH 7.2). Repeated thawing and freezing should be avoided. Working dilutions should be stored at +4°C, not refrozen, and preferably used the same day. If a slight precipitation occurs upon storage, this should be removed by centrifugation. It will not affect the performance or the concentration of the product.
CautionThis product is intended FOR RESEARCH USE ONLY, and FOR TESTS IN VITRO, not for use in diagnostic or therapeutic procedures involving humans or animals. It may contain hazardous ingredients. Please refer to the Safety Data Sheets (SDS) for additional information and proper handling procedures. Dispose product remainders according to local regulations.This datasheet is as accurate as reasonably achievable, but Exalpha Biologicals accepts no liability for any inaccuracies or omissions in this information.
References1. Roebroek, A.J.M., van de Velde, H.J.K., Van Bokhoven, A., Broers, J.L.V., Ramaekers, F.C.S., Van de Ven, W.J.M. (1993). Cloning and expression of alternative transcripts of a noval neuroendocrine-specific gene and identifiCation of its 135-kDa translation product. J. Biol. Chem. 268,13439-47. 2. van de Velde, H. J., Roebroek, A. J., van Leeuwen, F. W., and Van de Ven, W. J. (1994). Molecular analysis of expression in Rat brain of NSP-A, a novel neuroendocrine-specific protein of the endoplasmic reticulum, Mol Brain Res 23, 81-92. 3. van de Velde, H. J., Roebroek, A. J., Senden, N. H., Ramaekers, F. C., and Van de Ven, W. J. (1994). NSP-encoded reticulons, neuroendocrine proteins of a novel gene family associated with membranes of the endoplasmic reticulum, J Cell Sci 107, 2403-16. 4. van de Velde, H. J., Senden, N. H., Roskams, T. A., Broers, J. L., Ramaekers, F. C., Roebroek, A. J., and Van de Ven, W. J. (1994). NSP-encoded reticulons are neuroendocrine markers of a novel Category in Human lung cancer diagnosis, Cancer Res 54, 4769-76. 5. Senden, N. H., Timmer, E. D., de Bruine, A., Wagenaar, S. S., Van de Velde, H. J., Roebroek, A. J., Van de Ven, W. J., Broers, J. L., and Ramaekers, F. C. (1997). A comparison of NSP-reticulons with conventional neuroendocrine markers in immunophenotyping of lung cancers, J Pathol 182, 13-21 6. Hens, J., Nuydens, R., Geerts, H., Senden, N. H., Van de Ven, W. J., Roebroek, A. J., van de Velde, H. J., Ramaekers, F. C., and Broers, J. L. (1998). Neuronal differentiation is accompanied by NSP-C expression, Cell Tissue Res 292, 229-37.
CE MarkCE
Safety Datasheet(s) for this product:EA_Sodium Azide/wp-content/uploads/SDS/Antibody SDS with Sodium AzideV2.pdf
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1.标准曲线的标准品是否一定要梯度稀释,为什么?我试过非梯度稀释的,也可以达到线性R2=0.99.
2.我用了CurveExpert做标曲,自动搜索后发现有10种提供的方程,各种形式的,其中一个十分适合我的实验结果(LogisticModel),而其他的感觉又不适合,因为结果常常为负值。这又是为啥捏?
3.实验的酶标仪最大OD值可以测到4,如果我的测量结果在1.3,是否像其他人所说的>1了就不准确了。
4.利用夹心法进行定量分析是否一定要使用线性方程?
不好意思啊,一下问了这么多问题,最近做了一个月的ELISA,完全摸不清头脑啊。谢谢各位了
我现在用商品化的试剂盒进行夹心法ELISA测定某抗体的浓度,盒子给定了4个标准品,标准品抗体浓度为70000~1000单位,测定步骤中要求对样品进行1:1000稀释后测定。
但是有的样本按照1:1000稀释后,最终OD值大于70000浓度的标准品的OD值,然后使用ELISACALC软件进行四参数拟合,超过了标准曲线的范围,就算不出来这个样品的浓度,但是如果在1000-70000之间的OD值就可以算出相应浓度。请教大神们,接下来我是否可以对样品进行1:2000,1:4000浓度稀释,算出结果后再×2、×4,算作此样品的浓度呢?还是直接给一个>70000的定性结果?或者可以有能算出来的其他软件?
Ps试剂盒中提及Dilutionlinearity(稀释线性?)为141%,这个是什么意思呢?
多谢!!
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请大家帮帮忙,第一步加完抗原后必须封闭吗
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