StemLight? Pluripotency Antibody Kit

• 产品编号：CST-9656S      品牌：CST       原厂货号：9656S
• 产品分类：生化和细胞分析 > 其它细胞分析试剂盒 > 其它细胞分析试剂盒 > 其它
• 应用分类：

描述：

StemLight™多潜能抗体试剂盒中的各种抗体可分别识别各自内源性多潜能标志蛋白。此Nanog抗体由合成肽段免疫动物产生，合成的肽段与人源nanog蛋白氨基末端氨基酸序列一致。抗体由蛋白质A和肽段亲和层析纯化得到。Oct-4A抗体由与人源Oct-4A 蛋白氨基末端氨基酸序列一致的重组蛋白免疫动物产生。Sox2抗体由合成肽段免疫动物产生，合成的肽段与人源Sox2蛋白氨基末端的氨基酸序列一致。TRA-1-81, 和 TRA-1-60(S)抗体由人源胚胎性癌2102Ep cl.2A6细胞免疫动物产生。StemLight™多潜能抗体试剂盒包含一组抗体用于检测人类多潜能细胞特异表达的蛋白。此试剂盒可用于追踪人胚胎干细胞（ES）或诱导多潜能细胞（iPS）的多向潜能性。标志物的丧失暗示多分化潜能性的丧失或者培养细胞的分化。试剂盒中组分已经优化使适用于在免疫荧光分析，试剂足够进行50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：

Source / Purification

Nanog Antibody was produced by immunizing animals with a synthetic peptide corresponding to amino acid sequence at the amino terminus of human nanog. Antibodies are purified by Protein A and peptide affinity chromatography. Oct-4A antibody is produced by immunizing animals with recombinant protein specific to the amino terminus of human Oct-4A. Sox2 antibody is produced by immunizing animals with a synthetic peptide corresponding to amino acid sequences at the amino terminus of human Sox2. SSEA4, TRA-1-81, and TRA-1-60(S) antibodies are produced by immunizing animals with human embryonal carcinoma 2102Ep cl.2A6 cells.

Description

The StemLight™ Pluripotency Anitbody Kit contains a panel of antibodies for the detection of proteins that are specifically expressed in human pluripotent cells. The kit can be used to track the pluripotent potential of human embryonic stem (ES) or induced pluripotent (iPS) cells. The loss of these markers indicates a loss of pluripotency or differentiation of the culture. The kit components are pre-optimized for parallel use in immunofluorescent analysis. Enough reagents are provided for 50 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 antibody