Human CD272 (BTLA, B and T Lymphocyte Attenuator) is a member of the Immunoglobulin superfamily and has sequence homology to PD-1 and CTLA4. It is expressed on T and B lymphocytes and other hemopoetic lineages. Engagement of this molecule by its ligand, CD270(HVEM, a TNF superfamily member) can down regulate activated T and B cell responses. CD272 levels on antigen specific CD8+ T cells have been reported to decrease in viral specific, but not melanoma specific activated lines (1). Polymorphism of the CD272 molecule has been linked to rheumatoid arthritis (3).
Antibodies ANC6E9 and ANC5A5(cat #372-020) each bind to a distinct epitope of CD272 and are suitable as a matched pair for EIA (see attachment).
Isotype: Murine IgG1k
Immunogen: Recombinant extracellular human CD272
Specificity: Antibody from clone ANC6E9 binds to CD272 on the cell surface of human PBMC in Flow cytometry, and to recombinant CD272-muIg (cat #542-020) in EIA.
Functional Application: Antibody ANC6E9 blocks binding of Biotinylated CD270(HVEM)-muIg(cat #531-030) to CD272-muIg in EIA.
References:
1) Derre’ L, DE Speiser, et al. (2010) J Clin Invest 120(1):157-167.
2) Otsuki N, Azuma M, et al. (2006) Biochem Biophys Res Com 344(4): 1121-1127.
3) Lin SC, Chan CH, et al. (2006) J Biomed Sci 13(6): 853-860.
4) Pasero C, D Olive, et al. (2009) Curr Mol Med 9(7): 911-927.
5) Gavrieli M, KM Murphy, et al. (2006) Adv Immunol 92: 152-185.
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想用RNAiMAX或者lipo2000转染siRNA，但是看了下转染试剂的说明书和锐博的siRNA说明书，觉得分别对siRNA的用量描述差别挺大的呀，不知道到底该参考哪个呢。
以24孔板为例在siRNA说明书中写到，siRNA终浓度是50nM的话，加入浓度为20μM的siRNA1.25ul，每孔体积是500ul，那这样的话，每孔最终siRNA的量是25pmol。
但是在RNAiMAX或者是lipo2000说明书中，一个写的每孔siRNA用量是5pmol，一个是500ng，这与siRNA厂家所提供的量相差也太多了吧。
到底该看哪一个呢。
ps.一旦siRNA的量和体积确定下来之后，转染试剂的量和siRNA1:1的加就可以了吗？
请各位大神解答。
1.siRNA说明书中的用量，红线圈出
2.RNAIMAX说明书中siRNA的用量。
3.lipo2000说明书中siRNA用量
有战友对比过罗氏X-tremeGENEDNATransfectionReagent和lipofectamine3000的优劣吗？求解答
EntransterTM-R4000是英格恩生物（EngreenBiosystem）最新研发合成的针对siRNA、microRNA、mimic、inhibitor、mRNA和shRNA等RNA的转染试剂。EntransterTM-R4000不仅可以转染小RNA，而且针对mRNA等长链RNA优化。由于纳米技术的应用，EntransterTM-R4000能做到高达90%的转染效率和60-90%的沉默效率，对难转染细胞和原代细胞也非常适用。
先附上一篇英格恩生物RNA转染试剂转染mir-346至Jurkat细胞研究甲状腺的文献，以供参考。
RNA转染试剂转染Mir-346至Jurkat细胞研究甲状腺.pdf(5947.72k)
