Studying protein-protein interactions via blot overlay or Far Western blot.

نویسنده

  • Randy A Hall
چکیده

Blot overlay is a useful method for studying protein-protein interactions. This technique involves fractionating proteins on SDS-PAGE, blotting to nitrocellulose or PVDF membrane, and then incubating with a probe of interest. The probe is typically a protein that is radiolabeled, biotinylated, or simply visualized with a specific antibody. When the probe is visualized via antibody detection, this technique is often referred to as "Far Western blot." Many different kinds of protein-protein interactions can be studied via blot overlay, and the method is applicable to screens for unknown protein-protein interactions as well as to the detailed characterization of known interactions.

برای دانلود رایگان متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Production and characterization of polyclonal antibody against a synthetic peptide from β-actin protein

Objective(s):Antibodies against actin, as one of the most widely studied structural and multifunctional housekeeping proteins in eukaryotic cells, are used as internal loading controls in western blot analyses. The aim of this study was to produce polyclonal antibody against a synthetic peptide derived from N-terminal region of β-actin protein to be used as a protein loading control in western ...

متن کامل

Development of a recombinant protein-based dot-blot hybridization assay for the detection of antibody to chicken infectious bronchitis virus

Nucleocapsid (N) protein of infectious bronchitis virus (IBV), one of the viral structural proteins, inducesstrong antibody response in natural infection. In this study, a simple, recombinant N protein-based dot-blottest was developed to serologically examine chicken serum samples for the presence of IBV antibody.Initially, 72 serum samples were tested for the presence of IBV antibody using a c...

متن کامل

Extraction, Cloning and Expression of RTB, as a Vaccine Adjuvant/Carrier,in E. coli and Production of Mouse Polyclonal Antibody (Anti-B chain Abs)

        Ricin, the toxic lectin extracted from the castor bean plant (Ricinus communis), consists of an A chain (RTA) and a B chain (RTB). Anti-A chain Abs and anti-B chain Abs can neutralize toxins in vivo and in vitro via blocking the binding of the toxin to the cell. Also, RTB protein is able to serve as an antigen deliver to the mucosal immune system and act as an immunoad...

متن کامل

P-12: Direct Evaluation of HspA2 in Fertile and Infertile Individuals

Background: Heat-shock protein A2 (HspA2) is a testis-specific member of the HSP70 family known to play a critical role in spermatogenic cell differentiation. HSPA2 is correlated with sperm maturity, function and fertility, and diminished expression of HSPA2 results in abnormal sperm maturity and infertility. The aim of this study was to assess maturity of ejaculated spermatozoa via comparing p...

متن کامل

Immunofluorescent protein detection in Western blotting

This report describes the detailed procedures for Western blot analysis using fluorescent antibodies. After electrophoresis and subsequent electroblotting, the fluorescent-labeled antibodies were visible upon ultraviolet illumination of the polyvinylidene fluoride (PVDF) membranes and could then be photographed to give an accurate record of the blots. Fluorescent labeling allows for photographi...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

عنوان ژورنال:
  • Methods in molecular biology

دوره 261  شماره 

صفحات  -

تاریخ انتشار 2004