Microplate capture and detection assay is used to detect the specific protein-DNA interactions and identifies the proteins. The method includes immobilization of DNA probes on the surface of microplates (having wells) coated with streptavidin. The cellular extract, prepared in binding buffer, is added for enough time to facilitate binding protein to bind to the oligonucleotide followed by washing of wells several times to remove nonspecifically bound proteins. A protein specific antibody is used for detection of protein. Comparison of advantages and disadvantages of microplate capture and detection assay and EMSA: A: Advantages 1: Microplate capture and detection assay is highly sensitive, fast and throughput technique that is based on ELISA while EMSA is based on comparative velocity of migration of DNA-protein complexes through non denaturing gel electrophoresis. 2: Both techniques can skip the use of radioacitve probes by using biotinylated or fluorescently labeled DNA probes. 3: EMSA uses DNA probe mutational analysis for testing the binding affinity of proteins. 4: Though EMSA can detect the presence of low quantities of DNA binding proteins from lysates, Microplate capture and detection assay can detect even lesser abundance of protein in the well by using with enzyme-labeled antibodies and a chemiluminescent substrate. Disadvantages: 1: Both techniques require supershift antibodies i.e. antibodies which can selectively bind to DNA bound native proteins. 2: The technique gives little information about corresponding changes in transcription factor- DNA. 3: The techniques have been used for a few target proteins so far while EMSA is applicable under in vitro conditions only. Solution Microplate capture and detection assay is used to detect the specific protein-DNA interactions and identifies the proteins. The method includes immobilization of DNA probes on the surface of microplates (having wells) coated with streptavidin. The cellular extract, prepared in binding buffer, is added for enough time to facilitate binding protein to bind to the oligonucleotide followed by washing of wells several times to remove nonspecifically bound proteins. A protein specific antibody is used for detection of protein. Comparison of advantages and disadvantages of microplate capture and detection assay and EMSA: A: Advantages 1: Microplate capture and detection assay is highly sensitive, fast and throughput technique that is based on ELISA while EMSA is based on comparative velocity of migration of DNA-protein complexes through non denaturing gel electrophoresis. 2: Both techniques can skip the use of radioacitve probes by using biotinylated or fluorescently labeled DNA probes. 3: EMSA uses DNA probe mutational analysis for testing the binding affinity of proteins. 4: Though EMSA can detect the presence of low quantities of DNA binding proteins from lysates, Microplate capture and detection assay can detect even lesser abundance of protein in the well by us.

1. Microplate capture and detection assay is used to detect the specific protein-DNA interactions
and identifies the proteins. The method includes immobilization of DNA probes on the surface of
microplates (having wells) coated with streptavidin. The cellular extract, prepared in binding
buffer, is added for enough time to facilitate binding protein to bind to the oligonucleotide
followed by washing of wells several times to remove nonspecifically bound proteins. A protein
specific antibody is used for detection of protein.
Comparison of advantages and disadvantages of microplate capture and detection assay and
EMSA:
A: Advantages
1: Microplate capture and detection assay is highly sensitive, fast and throughput technique that
is based on ELISA while EMSA is based on comparative velocity of migration of DNA-protein
complexes through non denaturing gel electrophoresis.
2: Both techniques can skip the use of radioacitve probes by using biotinylated or fluorescently
labeled DNA probes.
3: EMSA uses DNA probe mutational analysis for testing the binding affinity of proteins.
4: Though EMSA can detect the presence of low quantities of DNA binding proteins from
lysates, Microplate capture and detection assay can detect even lesser abundance of protein in the
well by using with enzyme-labeled antibodies and a chemiluminescent substrate.
Disadvantages:
1: Both techniques require supershift antibodies i.e. antibodies which can selectively bind to
DNA bound native proteins.
2: The technique gives little information about corresponding changes in transcription factor-
DNA.
3: The techniques have been used for a few target proteins so far while EMSA is applicable
under in vitro conditions only.
Solution
Microplate capture and detection assay is used to detect the specific protein-DNA interactions
and identifies the proteins. The method includes immobilization of DNA probes on the surface of
microplates (having wells) coated with streptavidin. The cellular extract, prepared in binding
buffer, is added for enough time to facilitate binding protein to bind to the oligonucleotide
followed by washing of wells several times to remove nonspecifically bound proteins. A protein
specific antibody is used for detection of protein.
Comparison of advantages and disadvantages of microplate capture and detection assay and

2. EMSA:
A: Advantages
1: Microplate capture and detection assay is highly sensitive, fast and throughput technique that
is based on ELISA while EMSA is based on comparative velocity of migration of DNA-protein
complexes through non denaturing gel electrophoresis.
2: Both techniques can skip the use of radioacitve probes by using biotinylated or fluorescently
labeled DNA probes.
3: EMSA uses DNA probe mutational analysis for testing the binding affinity of proteins.
4: Though EMSA can detect the presence of low quantities of DNA binding proteins from
lysates, Microplate capture and detection assay can detect even lesser abundance of protein in the
well by using with enzyme-labeled antibodies and a chemiluminescent substrate.
Disadvantages:
1: Both techniques require supershift antibodies i.e. antibodies which can selectively bind to
DNA bound native proteins.
2: The technique gives little information about corresponding changes in transcription factor-
DNA.
3: The techniques have been used for a few target proteins so far while EMSA is applicable
under in vitro conditions only.