Boster Bio Life Science Blog

What is a western blot?

Western blotting (also called Protein Immunoblotting) is an analytical technique used to detect specific proteins in the given sample. It uses SDS-polyacrylamide gel electrophoresis (SDS-PAGE) to separate various proteins contained in the sample. The separated proteins are then transferred or blotted onto a matrix, where they are stained with antibodies specific to the target protein. Expression details of the target proteins in the given cells or tissue homogenate can then be obtained through analyzing the location and intensity of the specific reaction. Western blotting analysis can detect target protein as low as 1 ng due to high resolution of the gel electrophoresis and strong specificity and high sensitivity of the immunoassay. This method is used in the fields of molecular biology, biochemistry, immunogenetics and other molecular biology disciplines for various experiments.

What is Enhanced Chemiluminescence Western Blot Substrate?

Enhanced Chemiluminescence (ECL) Western Blot Substrate is a very sensitive, non-radioactive, enhanced luminol-based chemiluminescent substrate that allows for easy detection of horseradish peroxidase (HRP) on immunoblots. HRP is a common molecule conjugated to antibodies. ECL Western Blot Substrate has the capability of offering...

What is a western blot?

Western blotting (also called Protein Immunoblotting) is an analytical technique used to detect specific proteins in the given sample. It uses SDS-polyacrylamide gel electrophoresis (SDS-PAGE) to separate various proteins contained in the sample. The separated proteins are then transferred or blotted onto a matrix, where they are stained with antibodies specific to the target protein. Expression details of the target proteins in the given cells or tissue homogenate can then be obtained through analyzing the location and intensity of the specific reaction. Western blotting analysis can detect target protein as low as 1 ng due to high resolution of the gel electrophoresis and strong specificity and high sensitivity of the immunoassay. This method is used in the fields of molecular biology, biochemistry, immunogenetics and other molecular biology disciplines for various experiments.

What is western blot filter paper?

Filter paper (or blotting paper) is important to ensure quick and efficient transfer of molecules from the electrophoresis gel to the matrix membrane. It is used for transfer sandwiches and cassettes during Western blotting and assists with transferring proteins unto polyvinylidene fluoride (PVDF), nitrocellulose, and other types of membranes. Western blot filter paper is a semi-permeable paper barrier used to separate fine solid particles from liquids...

What is a nitrocellulose membrane?

Nitrocellulose membranes are one of the top matrices used in protein blotting. They have high protein-binding affinity, compatibility with a variety of detection methods, and the ability to immobilize proteins, glycoproteins, or nucleic acids. Examples of compatible detection methods include chemiluminescence, chromogenic, and fluorescence. It is proven to produce excellent signal-to-noise results when used for amino acid analysis and western, northern, and Southern blotting.

What is a western blot?

Western blotting (also called Protein Immunoblotting) is an analytical technique used to detect specific proteins in the given sample. It uses SDS-polyacrylamide gel electrophoresis (SDS-PAGE) to separate various proteins contained in the sample. The separated proteins are then transferred or blotted onto a matrix, where they are stained with antibodies specific to the target protein. Expression details of the target proteins in the given cells or tissue homogenate...

ELISA (enzyme-linked immunosorbent assay) is a plate-based assay used to detect the concentration of a specific protein in a liquid sample. Three different types of data output can be obtained:

Types of Data Output

• Qualitative: ELISAs can be used to determine whether or not the protein of interest is present in the sample by comparing the sample to a blank well or an unrelated control antigen without the target protein.
• Semi-Quantitative: Since the signal intensity produced is directly related to the amount of target protein in the sample, ELISA allows samples to be compared by observing the relative levels of the target protein.
• Quantitative: Quantitative: For a precise calculation of the target protein concentration in assay samples, a standard curve with known target protein concentrations should be created using a purified antigen and compared with the ELISA data generated.

To set up a standard curve, ELISA standards should be carefully prepared for accurate sample quantification of...

ELISA (enzyme-linked immunosorbent assay) is a convenient and simple method to quantitatively or qualitatively detect peptides, proteins, antibodies, and hormones in samples, rendering it as one of the most widely used immunoassays. Despite the many advantages of conducting ELISA, there are some mistakes that could turn your ELISA experiment sour. Help prevent this situation from happening by avoiding 5 common pitfalls when performing an ELISA:

TMB Coloring Development Agent Dilemma

1. “I think I still have some of the TMB coloring development agent from the previous kit. Maybe I’ll use that instead.”

Stop! Wanting to conserve resources is a good habit, but not in this case. We should avoid using reagents from different batches together. Each...

The ELISA (enzyme-linked immunosorbent assay) is recognized by scientists for its many advantages. The assay is convenient, quick, and simple to execute. ELISA’s versatility to detect peptides, proteins, antibodies, and hormones, and its ability to generate quantitative and qualitative data make it one of the most popular and powerful immunoassays available.

In response to popular demand, numerous commercial ELISA kits are offered in the market, but not all ELISA kits are created equal.

How do we sift through the masses and choose a good ELISA kit?

Keep in mind the following points next time you go hunting for an ELISA kit.

Assay Format

• What would you like to determine for your sample? There are four categories of ELISAs - direct, indirect, sandwich, and competitive.
• If you’re planning for speedy detection of immune response to an antigen, and signal amplification isn’t a priority, direct ELISA might be for you.
• For better sensitivity and greater flexibility to figure out the total antibody concentration, indirect ELISA might be more suitable.
• What if your sample is complex? Fear not, choose sandwich ELISA for high sensitivity, specificity, and f
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