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Western Blot Services

KMD Bioscience has been working on protein expression research for over 10 years. Our scientists, which have accumulated a wealth of experience in experimental operations, are able to quickly complete the specified experiments in a relatively short period of time, while ensuring that the test results are accurate, objective and credible. KMD Bioscience has established a perfect protein platform with sophisticated protein detection equipment and technology, which can provide one-stop technical services from recombinant protein preparation to protein expression and purification, protein function research and other one-stop technical services to ensure high-quality service content. We have the ability to provide customers with Western Blot and other commonly used immunological detection services.

Western Blot, is one of the common technical methods for qualitative and quantitative detection of protein expression. The primary aim of Western Blot is to detect if a certain protein (target protein) is expressed in a sample (tissue or cell) and the relative abundance of expression. Through the specific recognition and binding of antibodies to the target proteins in gel electrophoresis-treated cells or biological tissue samples, the expression of specific protein levels in cells or tissues can be analyzed according to the location and depth of coloration. This technology is a new immunobiochemical technology developed on the basis of gel electrophoresis and solid-phase immunoassay technology, so it has both the high resolution and high specificity of SDS-PAGE and solid-phase immunoassay technology, which plays an important role in proteomics as a convenient and reliable research tool, and mass spectrometry and protein chip technology.

Western Blot experimental services have developed to this day and are widely used. But getting a clear background, bright strips, real and objective experimental results is not easy, and it takes a lot of time and effort. KMD Bioscience has the skills to summarize the details of experimental operations, has strict quality control points, rigorous pre-experimental settings, can tailor different experimental protocols for different customers' samples, and provide clear and complete Western Blot experimental results, saving customers' valuable scientific research time and meeting customer needs with high quality.

 

Western Blot

Experimental principle

The technique is based on the principle of specific binding of antigen and antibody to detect a protein in complex samples. Proteins in cell or tissue samples are separated by polyacrylamide gel electrophoresis, and after membrane transfer and sealing, the target protein binds to a specific primary antibody, which reacts with a horseradish peroxidase (HRP)-labeled secondary antibody, and then undergoes substrate chromatography, which analyzes the position of the exposed bands and gray scale analysis to determine the expression of the target protein in the sample.

Applications

*Widely used for protein expression level detection.

*Identification of a protein, also quantitative and semi-quantitative analysis of the target protein.

*Can be used for subsequent analysis of protein-protein, protein-DNA and protein-RNA interactions.

Advantages

*The required equipment is convenient, low cost and flexible to use.

*Simple operation, generally stable results, and positive results are generally more reliable.

*Results analysis is easier than mass spectrometry.

 

The Main Process of the Western Blot Experiment:

 

 

 

With the upgrading of electrophoresis and electrotransfer equipment, the commercialization of antibodies and signal detection systems, and other technological advances, the sensitivity of Western Blot has been improved, and Western Blot is easy to operate and therefore can be performed in general laboratories, but a variety of problems are often encountered in the process of experimentation.

 

Frequently Occurring Problems and Countermeasures :

 

Experimental Problems

Possible Reasons

Recommendations

Destination strip not visible

Expression of the target protein is too low

Increased sample volume, concentration of target proteins, use of higher sensitivity kits

Insufficient potency of antibodies used

Use freshly prepared primary antibody, avoid repeated freezing and thawing; increase the concentration of primary or secondary antibody and prolong the incubation time appropriately

Low efficiency of membrane transfer

Selection of the appropriate film transfer method to improve film transfer efficiency

The strips are non-parallel and curved

Electrophoretic migration too fast or migration temperature too high

Adjust electrophoresis settings such as PH, voltage, etc., and lower the ambient temperature during electrophoresis

The background is very high

Inappropriate closure conditions

Extend the sealing time or change the sealer

High antibody concentration

Reduced primary/secondary antibody concentration and extended incubation time

Antibodies cross-react with sealers

Add heat and a stain remover such as Tween 20

 

Service Advantages:

 

-- For different protein samples, Kamed Bio adopts different protein concentration determination methods (Lowry method or BCA method)

-- Assurance of the quality of the experiment, strict QC quality control: the use of internal reference for accurate quantification, the same sample, protein concentration quantitative value of the difference between several experiments is less than 8%

-- Reproducibility between different personnel: to ensure the accuracy of the results

-- Complete technical services, we are able to provide technical services from gene synthesis, cell transfection, vector construction, protein expression and purification, protein function research, etc

-- Many years of experience in antibody research, with a wealth of knowledge on antibody selection, to avoid false-positive results from the root causes

 

How to Order?

 

If you have any questions regarding our services or products, please feel free to contact us by E-mail: info@kmdbioscience.com or Tel: +86-022-8216-4980;