Intermediate Filaments Antibody Sampler Kit

• 产品编号：CST-4751S      品牌：Cell Signaling Technology       原厂货号：4751S
• 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：

Pan-Keratin (C11) Mouse mAb检测内源性keratin 4、5、6、8、10、13和18总蛋白，但是不能与其它keratin蛋白发生交叉反应。Plectin-1 Antibody检测内源性plectin-1总蛋白，并且期望基于免疫原性多肽的序列能够与plectin-1蛋白所有的亚型发生交叉反应。GFAP (GA5) Mouse mAb检测内源性GFAP总蛋白。Desmin Antibody检测内源性desmin总蛋白。Vimentin (R28) Antibody检测内源性vimentin总蛋白。通过来自A431细胞的一个细胞骨架的制备去免疫动物从而生成出Pan-Keratin (C11) monoclonal antibody。通过来自猪脊髓的天然纯化的GFAP蛋白去免疫动物从而制备出GFAP (GA5) monoclonal antibody。通过人工合成人源desmin蛋白羧基末端相应的多肽片段去免疫动物从而制备出Desmin monoclonal antibody。通过人工合成人源plectin-1蛋白中间序列和人源vimentin蛋白Arg28位点周围相应的多肽片段去免疫动物从而制备出多克隆抗体。通过多肽亲和层析纯化抗体。该Intermediate Filaments Antibody Sampler Kit提供一个经济手段去评价中间丝的存在和水平。该试剂盒包含充足的一抗和二抗，每个抗体能进行四次免疫印迹（Western blot）实验。细胞骨架由三种细胞基质纤维组成：微丝(actin filaments)、中间纤维和微管。中间纤维的主要类型是有区别的，且它在特别的细胞中表达：细胞角蛋白（epithelial细胞）、胶质原纤维酸性蛋白 或GFAP（glial细胞）、desmin （skeletal、viscera和某种vascular smooth muscle 细胞）、波形蛋白（间充质来源）和神经纤维细丝（neurons）。GFAP和vimentin蛋白在星形胶质细胞瘤细胞中形成中间纤维，并且调节它们的活性和形状(1)。尤其，vimentin filaments在早期发育阶段出现，而GFAP filaments是分化的和成熟的大脑星形胶质细胞的特征。因此，GFAP普遍被用作来自星形胶质细胞颅内和椎管内肿瘤上升的标记物 (2)。Vimentin出现在恶性毒瘤中，但不出现癌中，因此相对于其它标记物该蛋白的表达被检测用于区别两种形式的肿瘤(3) 。Desmin蛋白是肌原性的标记物，其表达在早期filaments网络形成的发展期，其恩呢该延伸穿过肌原纤维和Z discs周围。desmin细胞骨架提供了一个在肌原纤维、细胞器和细胞骨架之中的连接(4)。Desmin 基因敲除的小鼠会发展成心肌病、骨骼肌和平滑肌缺陷(5)。在人类，通过在相应的desmin基因或在desmin 相互作用的蛋白包括αB-crystallin 和synemin蛋白的突变，desmin蛋白相关的肌肉疾病可能被引起。紊乱的desmin filaments和蛋白质聚合物主要包含desmin的堆积都是desmin相关的肌肉疾病的特征 (6,7)。Keratins蛋白能装配成中间纤维，其形成了酸性角蛋白（I型，角蛋白9-23）和中性或碱性角蛋白（II型，角蛋白1-8）的异源二聚体(8,9)。Keratin蛋白亚型证明组织和分化特异性的特点，这使它们用作生物标记物(8)。keratin基因突变与皮肤病、肝脏和胰腺疾病和炎性肠道疾病有关联(10-13)。Plectin蛋白是一个广泛表达的蛋白，该蛋白与中间丝和肌动蛋白细胞骨架、机械稳定的细胞和组织有交联。Plectin蛋白也在调节肌动蛋白动力中起着一定作用，并且该蛋白用作信号分子的支架蛋白(14)。Plectin蛋白在半桥粒的稳定性有着重要作用，并且交联它们到中间丝的网架中。研究已经显示Plectin蛋白涉及数个信号传导通路。通过结合和隔绝RACK1激酶、activated C kinase 1 受体，它能传递信号到PKC激酶(15,16)。Plectin蛋白也涉及角蛋白架构和细胞应激反应(16)的调节、通过趋化因子受体CXCR4蛋白信号的传递(17)、AMP-activated protein kinase (AMPK)活性的调节和小鼠肌管的信号传递(18)。

原厂资料：

Specificity / Sensitivity

Pan-Keratin (C11) Mouse mAb detects endogenous levels of total keratin 4, 5, 6, 8, 10, 13 and 18 and does not cross-react with other keratins. Plectin-1 Antibody detects endogenous levels of total plectin-1 protein and is expected to cross-react with all isoforms of plectin-1 based on the sequence of the immunogenic peptide. GFAP (GA5) Mouse mAb detects endogenous levels of total GFAP protein. Desmin Antibody detects endogenous levels of total desmin protein. Vimentin (R28) Antibody detects endogenous levels of total vimentin protein.

Source / Purification

Pan-Keratin (C11) monoclonal antibody is produced by immunizing animals with a cytoskeleton preparation from A431 cells. GFAP (GA5) monoclonal antibody is produced by immunizing animals with native GFAP purified from pig spinal cord. Desmin monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to carboxy terminal residues of human desmin protein. Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to the central sequence of human plectin-1 and residues surrounding Arg28 of human vimentin. Antibodies are purified by peptide affinity chromatography.

Background

The cytoskeleton consists of three types of cytosolic fibers: microfilaments (actin filaments), intermediate filaments and microtubules. Major types of intermediate filaments are distinguished and expressed in particular cell types: cytokeratins (epithelial cells), glial fibrillary acidic protein, GFAP (glial cells), desmin (skeletal, visceral and certain vascular smooth muscle cells), vimentin (mesenchyme origin) and neurofilaments (neurons). GFAP and vimentin form intermediate filaments in astroglial cells and modulate their motility and shape (1). In particular, vimentin filaments are present at early developmental stages, while GFAP filaments are characteristic of differentiated and mature brain astrocytes. Thus, GFAP is commonly used as a marker for intracranial and intraspinal tumors arising from astrocytes (2). Vimentin is present in sarcomas, but not carcinomas, and its expression is examined in conjunction with that of other markers to distinguish between the two (3). 
 Desmin is a myogenic marker expressed in early development that forms a network of filaments that extends across the myofibril and surrounds Z discs. The desmin cytoskeleton provides a connection between myofibrils, organelles and the cytoskeleton (4). Desmin knockout mice develop cardiomyopathy as well as skeletal and smooth muscle defects (5). In humans, desmin related myopathies might be caused by mutations in the corresponding desmin gene or in proteins with which desmin interacts, including αB-crystallin and synemin. Disorganized desmin filaments and the accumulation of protein aggregates comprised predominantly of desmin characterize desmin-related myopathies (reviewed in 6,7). 
 Keratins assemble into filaments, forming heterodimers of an acidic keratin (or type I keratin, keratins 9 to 23) and a basic keratin (or type II keratin, keratins 1 to 8) (8,9). Keratin isoforms demonstrate tissue- and differentiation-specific profiles, which make them useful as biomarkers (8). Mutations in keratin genes are associated with skin disorders, liver and pancreatic diseases, and inflammatory intestinal diseases (10-13). 
 Plectin is a large, widely expressed protein that crosslinks the intermediate filament and actin cytoskeleton, mechanically stabilizing cells and tissues. Plectin also plays a role in the regulation of actin dynamics and acts as a scaffold for signaling molecules (14). It is important in the stabilization of hemidesmosomes, crosslinking them to the intermediate filament network. Plectin has been shown to be involved in several signaling cascades. It signals to PKC by binding to and sequestering RACK1, the receptor for activated C kinase 1 (15,16). Plectin is also involved in the regulation of cytokeratin architecture and cell stress response (16), signaling through the chemokine receptor CXCR4 (17), regulation of AMP-activated protein kinase (AMPK) activity and signaling in mouse myotubes (18).

注意事项：

Storage: Supplied in 10 mM sodium HEPES (pH 7.5), 150
mM NaCl, 100 µg/ml BSA , 50% glycerol and less than 0.02%
sodium azide. Store at –20°C. Do not aliquot the antibodies