Ccn Proteins: Methods and Protocols

ISBN-13: 9781493964284 / Angielski / Twarda / 2016 / 576 str.

Masaharu Takigawa

Opis

This volume provides a comprehensive guide on the Cyr61 (Cysteine-rich 61) (CCN) family of proteins and genes from basic research to cutting-edge methodologies and state-of-the-art techniques. Chapters details practical tips to overcome any obstacles with experimentation pertaining to chemistry, biology, physiology, pathology, medical and dental sciences and pharmacology of CCN proteins. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, CCN Proteins: Methods and Protocols will be a valuable resource for a wide audience, ranging from the experienced CCN researchers looking for new approaches to junior graduate students just beginning in CCN research.

Spis treści

The CCN Proteins: An Overview.- Gene Expression Analysis of CCN Proteins: Whole Mount In Situ Hybridization Of CCN2 in Developing Calcified Tissues .- Expression of CCN Genes and Proteins in Human Skin: Methods And Protocols.- Analysis of Expression of CCN Family Genes in Skeletal Tissue-Derived Cells.- Western Blotting Analysis of CCN Proteins.-  Immunohistochemical Analysis of CCN Proteins.- Analysis of CCN Expression by Immunofluorescence on Skin Cells, Skin, and Reconstructed Epidermis.-  Production of Recombinant CCN2 Protein in Escherichia. Coli.-  Production Of Recombinant CCN Proteins By Brevibacillus Choshinensis.- Production Of Recombinant CCN2 Protein By Mammalian Cells.- In Vitro Transfection with and Expression of CCN Family of Genes.- Preparation of Module Specific Antibodies against CCN2.- ELISA of CCN Proteins in Body Fluids Including Serum And Plasma.- Analysis of Signaling Pathways Activated by CCN Proteins.- Protocols for Screening for Binding Partners of CCN Proteins: Yeast Two- Hybrid System.- Protocols For Screening Peptide Motifs Binding To CCN Proteins.- Evaluation Of Molecular Interaction Between CCN Proteins and its Binding Partners By Surface Plasmon Resonance (SPR).-  Promoter Analysis of CCN Genes.- Analysis Of Posttranscriptional Regulation Of CCN Genes.-  Protein Imaging of CCN2 and CCN3 in Living Cells.-  Cell Biological Assays for Measuring Chondrogenic Activities of CCN Proteins.-  Protocol for Measuring Angiogenic Activities of CCN Proteins.- Cell Biological Assays for Measuring Odontogenic Activities of CCN Proteins.- Separation and Enrichment of Haematopoietic Stem Cells for CCN Studies.- In Vivo Evaluation of Cartilage Regenerative Effects of CCN Proteins.- Gene Expression Analysis of CCN Proteins in Bone Under Mechanical Stress.- The Bone Regeneration Model and Primary Osteoblastic Cell Culture Used in the Analysis of CCN3 Transgenic and Knockout Mice.- Design and Analysis of CCN Gene Activity Using CCN Knockout Mice Containing Lacz Reporters.- Analysis of CCN4 Function in Osteogenic and Osteoclastic Cells Using Gain and Loss of Function Approaches.- Construction and Analysis of an Allelic Series of CCN1 Knockin Mice.-  Production and Analysis of Conditional KO Mice of CCN2 in Kidney.- Generation And Analysis Of Cartilage-Specific CCN2 Overexpression In Transgenic Mice.- Analysis of Transcytosis of CCN2 by Chondrocytes.- CCN2 in Skin Fibrosis.- Studying the CCN Proteins in Fibrosis.- Analysis of Pathological Activities of CCN Proteins in Fibrotic Diseases - Kidney Fibrosis.- Analysis of Pathological Activities of CCN Proteins in Fibrotic Diseases - Liver Fibrosis.- Cellular or Exosomal Micrornas Associated with CCN Gene Expression in Liver Fibrosis.- TGF-Β1 snd CCN2-Stimulated Sirius Red Assay for Collagen Accumulation in Cultured Cells.-  CCN Detection of Cancer Tissues by Immunohistochemistry Staining.- Gene Expression Analysis of CCN Proteins: Detection of CCN1 and CCN5 MRNA in Human Cancer Samples using a Modified In Situ Hybridization Technique.- Analysis of Pathological Activities of CCN Proteins in Bone Metastasis.- Analysis of Pathological Activities of CCN2/CTGF in Muscle Dystrophy.-  Method for Analysis of Matrix Degradation by CCN2 through the MMP/TIMP System.- An Analysis of Pathological Activities of CCN Proteins in Joint Disorders−Mechanical Stretch-Mediated CCN2 Expression in Cultured Meniscus Cells−.-  Analysis of CCN Protein Expression and Activities in Vasoproliferative Retinopathies.