Western Blottin
Bisho raj
MSc Microbiol
National co
science and te
,INTRODUCTION
Western blot is the analytical technique used in molecular
biology, immunogenetics, and other molecular biology to
detect specific proteins in a sample of tissue homogenate
or extract
It is also called immunoblotting because an
antibody is used to specifically detect its antigen
In this technique a mixture of proteins is
separated based on molecular weight, and thus by
type, through gel electrophoresis
The process of western blotting involves the
transfer of proteins separated by SDS PAGE into
an absorbent membrane W. Neal Burne
Between 1979 and 2019 "it has been mentioned in the titles, abstr
keywords of more than 400,000 PubMed-listed publications" and may st
most used protein-analytical technique
, INTRODUCTION
Western blotting or protein blotting depends on the specificity of in
between the protein of interest and the probe used for the detection of the p
Unlike Southern blotting that utilizes radio-labeled nucleic acid probes
blotting usually uses a second antibody tagged with an enzyme
Applications
• Identification of a specific protein in a complex mixture of proteins
• Estimation of the size of the protein as well as the amount of protein p
the mixture
• Demonstration of specific antibodies in the serum for diag
neurocysticercosis and tubercular meningitis.
• Confirmatory HIV-test to detect anti-HIV antibody in a human serum sa
• Detection of HSV infections
• Confirmatory test for Hepatitis B infection
Bisho raj
MSc Microbiol
National co
science and te
,INTRODUCTION
Western blot is the analytical technique used in molecular
biology, immunogenetics, and other molecular biology to
detect specific proteins in a sample of tissue homogenate
or extract
It is also called immunoblotting because an
antibody is used to specifically detect its antigen
In this technique a mixture of proteins is
separated based on molecular weight, and thus by
type, through gel electrophoresis
The process of western blotting involves the
transfer of proteins separated by SDS PAGE into
an absorbent membrane W. Neal Burne
Between 1979 and 2019 "it has been mentioned in the titles, abstr
keywords of more than 400,000 PubMed-listed publications" and may st
most used protein-analytical technique
, INTRODUCTION
Western blotting or protein blotting depends on the specificity of in
between the protein of interest and the probe used for the detection of the p
Unlike Southern blotting that utilizes radio-labeled nucleic acid probes
blotting usually uses a second antibody tagged with an enzyme
Applications
• Identification of a specific protein in a complex mixture of proteins
• Estimation of the size of the protein as well as the amount of protein p
the mixture
• Demonstration of specific antibodies in the serum for diag
neurocysticercosis and tubercular meningitis.
• Confirmatory HIV-test to detect anti-HIV antibody in a human serum sa
• Detection of HSV infections
• Confirmatory test for Hepatitis B infection
