StemLight? iPS Cell Reprogramming Antibody Kit

• 产品编号：CST-9092S      品牌：Cell Signaling Technology       原厂货号：9092S
• 产品分类：抗体 > 一抗 > 复合抗体试剂盒
• 应用分类：

其它组分：

描述：

Oct-4A (C30A3) Rabbit mAb可以检测内源性总Oct-4A 蛋白。Sox2 (D6D9) XP® Rabbit mAb能检测内源性总Sox2 蛋白。Nanog (D73G4) XP® Rabbit mAb可以检测内源性总nanog蛋白。LIN28A (D84C11) XP® Rabbit mAb可以检测内源性总LIN28A蛋白。c-Myc (D84C12) XP® Rabbit mAb可以检测内源性总c-Myc蛋白。本抗体不推荐用于检测Myc-标签融合蛋白的抗体（应使用Cell Signaling Technology cat. #2276 or #2278 产品）。KLF4抗体可以用于在western实验中检测内源性总KLF4蛋白，但不推荐在免疫荧光实验中用于检测外源表达的蛋白。本试剂盒中所有产品与人蛋白的反应性已经过验证，但是部分产品与其他物种也有交叉反应。请参阅www.cellsignal.com以获得产品中个别组分的反应特异性/敏感性信息。合成与人源Oct-4A的重组蛋白、与人源Sox2蛋白的Gly179，与邻近人源Nanog蛋白的氨基末端等邻近氨基酸残基序列一致的肽段，人LIN28A羧基端邻近氨基酸残基序列一致的肽段和c-My体细胞重编程诱导多能干细胞（iPS细胞）的多肽免疫动物获得多克隆抗体。抗体通过蛋白A和肽亲和层析纯化。StemLight™ iPS cell Reprogramming Antibody Kit包括一组抗体用于检测在体细胞重编程诱导多能干细胞（iPS细胞）过程中使用的各种蛋白。试剂盒可以用于跟踪转染，病毒转导和其他蛋白表达方法以后重编程因子的表达效率。试剂盒中组分已经优化为标准稀释度用于在免疫荧光分析中配套使用，但部分组分经证明可以用于其他实验——单个产品的使用建议请参阅其说明书。试剂足够进行160次反应体系为100ul的免疫荧光实验。具有分化潜能的细胞能够分化为三种胚层，包括内胚层，外胚层和中胚层细胞。胚胎干细胞，胚胎瘤细胞和诱导多能干细胞均具有多潜能性。Oct-4, Sox2 和Nanog是在多潜能干细胞中高表达的重要转录因子（1）。它们共同构成转录网络，保持细胞处于多潜能状态（2,3）。过表达Oct-4 和Sox2以及Klf4和 c- Myc能诱导小鼠和人的体细胞具备多潜能性，说明在维持细胞更新和多潜能性所需的转录网络中它们是关键的调控因子（4-5）。过表达Oct-4 ,Sox2,Nanog和Lin28也能邮到人体细胞具备多潜能性（6）。在多潜能干细胞分化过程中，Oct-4 ,Sox2和Nanog表达量下降。SSEA4, TRA-1-81 和 TRA-1-60抗体可以识别所有多潜能干细胞细胞表面表达的抗原。SSEA4识别一种糖脂糖类抗原（7）。TRA-1-60(S) 和TRA-1-81抗体能识别细胞标志蛋白的多个变种的不同蛋白聚糖表位（8）。这些表位对神经酰胺酶敏感分别具有抗性。随着多能干细胞的分化失去多潜能性，SSEA4, TRA-1-81 和TRA-1-60抗体与细胞表面标记的反应也将消失（9）。

1. Looijenga, L.H. et al. (2003) Cancer Res 63, 2244-50.
2. Pesce, M. and Schöler, H.R. (2001) Stem Cells 19, 271-8.
3. Pan, G. and Thomson, J.A. (2007) Cell Res 17, 42-9.
4. Takahashi, K. and Yamanaka, S. (2006) Cell 126, 663-76.
5. Okita, K. et al. (2007) Nature 448, 313-7.
6. Yu, J. et al. (2007) Science 318, 1917-20.
7. Henderson, J.K. et al. (2002) Stem Cells 20, 329-37.
8. Draper, J.S. et al. (2002) J Anat 200, 249-58.
9. Schopperle, W.M. and DeWolf, W.C. (2007) Stem Cells 25, 723-30.

原厂资料：

Specificity / Sensitivity

Oct-4A (C30A3) Rabbit mAb detects endogenous levels of total Oct-4A protein. Sox2 (D6D9) XP® Rabbit mAb detects endogenous levels of total Sox2 protein. Nanog (D73G4) XP® Rabbit mAb detects endogenous levels of total nanog protein. LIN28A (D84C11) XP® Rabbit mAb detects endogenous levels of total LIN28A protein. c-Myc (D84C12) XP® Rabbit mAb detects endogenous levels of total c-Myc protein. This antibody is not recommended for detection of Myc-tagged fusion proteins (use Cell Signaling Technology cat. #2276 or #2278). KLF4 Antibody detects endogenous levels of total KLF4 protein by western blot, but for immunofluorescence it is only recommended for detection of exogenously expressed KLF4 protein. All components of this kit have been validated for reactivity to human protein, however some components also have additional species cross reactivities. Please see www.cellsignal.com for additional specificity/sensitivity information for individual kit components.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with recombinant protein specific to the amino terminus of human Oct-4A, a synthetic peptide corresponding to residues surrounding Gly179 of human Sox2, a synthetic peptide corresponding to residues near the amino terminus of human nanog protein, a synthetic peptide corresponding to residues near the carboxy terminus of human LIN28A, and a synthetic peptide correspondingto residues near the amino terminus of c-Myc. Polyclonal antibodies are produced by immunizing animals with synthetic peptide corresponding to residues near the amino terminus of human KLF4. Polyclonal antibodies are purified by Protein A and peptide affinity chromatography.

Description

The StemLight™ iPS cell Reprogramming Antibody Kit contains a panel of antibodies for the detection of various proteins, combinations of which have been used to reprogram somatic cells to Induced Pluripotent Stem (iPS) cells. The kit can be used to track efficiency of expression of the reprogramming factors following transfection, viral transduction and other means of protein delivery. The kit components are pre-optimized for parallel use in immunofluorescent analysis at a standard dilution, but components are also validated for use in other applications --please refer to individual data sheet information for application specific recommendations. Enough reagents are provided for 160 immunofluorescent assays based on a working volume of 100 μl.

Background

Pluripotency is the ability of a cell to differentiate into cell types of the three germ layers, the endoderm, ectoderm and mesoderm. It is a property shared by embryonic stem cells, embryonic carcinoma and induced pluripotent cells. 
 Oct-4, Sox2 and Nanog are key transcriptional regulators that are highly expressed in pluripotent cells (1). Together they form a transcriptional network that maintains cells in a pluripotent state (2,3). Over-expression of Oct-4 and Sox2 along with Klf4 and c- Myc can induce pluripotency in both mouse and human somatic cells, highlighting their roles as key regulators of the transcrip- tional network necessary for renewal and pluripotency (4-5). It has also been demonstrated that overexpression of Oct-4, Sox2, Nanog and Lin28 can induce pluripotency in human somatic cells (6). Upon differentiation of pluripotent cultures, expression of Oct-4, Nanog and Sox2 is downregulated. 
 SSEA4, TRA-1-81 and TRA-1-60 antibodies recognize antigens expressed on the cell surface of all pluripotent cells. SSEA4 recognizes a glycolipid carbohydrate epitope (7). TRA-1-60(S) and TRA-1-81 antibodies recognize different proteoglycan epitopes on variants of the same protein, podocalyxin (8). These epitopes are neurominadase sensitive and resistant, respectively. Reactivity of SSEA4, TRA-1-81 and TRA-1-60 antibodies with their respective cell surface markers are lost upon differentiation of pluripotent cells, corresponding with a loss of pluripotent potential (9).

1. Looijenga, L.H. et al. (2003) Cancer Res 63, 2244-50.
2. Pesce, M. and Schöler, H.R. (2001) Stem Cells 19, 271-8.
3. Pan, G. and Thomson, J.A. (2007) Cell Res 17, 42-9.
4. Takahashi, K. and Yamanaka, S. (2006) Cell 126, 663-76.
5. Okita, K. et al. (2007) Nature 448, 313-7.
6. Yu, J. et al. (2007) Science 318, 1917-20.
7. Henderson, J.K. et al. (2002) Stem Cells 20, 329-37.
8. Draper, J.S. et al. (2002) J Anat 200, 249-58.
9. Schopperle, W.M. and DeWolf, W.C. (2007) Stem Cells 25, 723-30.

注意事项：

Storage: Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM
NaCl, 100 μg/ml BSA and 50% glycerol. Store at –20°C. Do not
aliquot the antibodies.