96-well Cell Adhesion Assay
(Collagen IV Coated, Fluorometric Assay)
Catalog Number: ECM-1022 96 assays
Storage: Upon arrival store the components at 4C.
Introduction: Due to its diverse nature and composition, the ECM can serve many functions, such as providing support and anchorage for cells, segregating tissues from one another, and regulating intercellular communication. The ECM regulates a cell"s dynamic behavior. In addition, it sequesters a wide range of cellular growth factors, and acts as a local depot for them. Changes in physiological conditions can trigger protease activities that cause local release of such depots. This allows the rapid and local growth factor-mediated activation of cellular functions, without de novo synthesis.
Formation of the extracellular matrix is essential for processes like growth, wound healing and fibrosis. An understanding of ECM structure and composition also helps in comprehending the complex dynamics of tumor invasion and metastasis in cancer biology as metastasis often involves the destruction of extracellular matrix by enzymes such as serine and Threonine proteases and Matrix metalloproteinase.
Biopioneer 96-well Cell Adhesion Microplate Assays are designed for the determination of the relative attachment of adherent cell lines to extracellular matrix proteins such as clooagen I, collagen IV, Firbronectin and vitronectin.
Principle of the assay
BioPioneer 96-welCell Adhesion Assays are designed for the determination of cell attachment to ECM proteins. After incubation followed by a brief wash step, attached cells are quantified with fluorescent dye. BSA-coated wells serve as a negative control, and poly-L-lysine-coated wells serve as a positive control for general attachment.
Materials Provided
1. Collagen IV Adhesion Plate (Part No. ECM-10112): One 96-well plate containing 72 Bovine Collagen IV coated wells , 8 BSA-coated wells and 8 poly-lysine coated wells(see layout below)
2. Fluorecent dye Solution (Part #: 10003): 50 ul in an amber vial
3. PBS
Materials Not Supplied
1. Cell culture medium
2. Serum free medium, such as DMEM containing 0.5% BSA, 2 mM CaCl2 and 2 mM MgCl2
4. 1XD PBS
5. 96-well microtiter plate
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般基克隆物体直接取基包括面几步骤
提取植物或者物基组DNA（试剂盒）————做PCR需要基扩增（顺利2搞定）——扩增基做酶切——基连接想插入载体面（顺利1-2）——鉴定克隆基
步骤都商品化试剂或者试剂盒 要思路清晰做起快 1-2周能完
没证实情况任何没依据推断都假想空想
现在小弟碰见一个难题，我想克隆一个基因，但是我在NCBI一检索，发现别人只发表了这个基因的DNA全长，但是，CDNA没有发表，也没有发表文章，想请问下，在不知道CDNA的情况下，请问我应该如何做这个基因的克隆？？？（全基因组也不知道,其他生物没有这种基因）
详见：http://zhidao.baidu.com/question/1831844001436785260.html?fr=iks&word=%B2%A1%B6%BE%B5%C4%B8%B4%D6%C6%D6%DC%C6%DA%B5%C4%B9%FD%B3%CC%3F&ie=gbk
