KMD Bioscience Co., Ltd.
400-621-6806
Service Line：+1-315-239-3085
Address：FL-4, Building A5, International Enterprise Community, Tianjin, China
Email：[email protected]
KMD Bioscience has been working on protein expression research for over 10 years. Our scientists, who have accumulated a wealth of experience in experimental operations, can quickly complete the specified experiments in a relatively short period 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 production to protein purification, protein function research, and other one-stop technical services to ensure high-quality service content. We can provide customers with Western Blot analysis 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 Blotting 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 immunochemical technology developed based on 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 test experimental services have developed to this day and are widely used. However getting a clear background, bright strips, and 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, and rigorous pre-experimental settings, can tailor different experimental protocols for different customers' samples, and provide clear and complete Western Blotting analysis experimental results, saving customers' valuable scientific research time and meeting customer needs with high quality.
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Western Blot |
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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 grayscale analysis to determine the expression of the target protein in the sample. |
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Applications |
*Widely used for protein expression level detection. *Identification of a protein, as well as quantitative and semi-quantitative analysis of the target protein. *Can be used for subsequent analysis of protein-protein, protein-DNA, and protein-RNA interactions. |
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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 Blotting 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
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Experimental Problems |
Possible Reasons |
Recommendations |
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The destination strip is not visible |
Expression of the target protein is too low |
Increased sample volume, concentration of target proteins, use of higher sensitivity kits |
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Insufficient potency of antibodies used |
Use freshly prepared primary antibodies, avoid repeated freezing and thawing; increase the concentration of primary or secondary antibodies, and prolong the incubation time appropriately. |
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Low efficiency of membrane transfer |
Selection of the appropriate film transfer method to improve film transfer efficiency |
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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 |
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The background is very high |
Inappropriate closure conditions |
Extend the sealing time or change the sealer |
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High antibody concentration |
Reduced primary/secondary antibody concentration and extended incubation time |
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Antibodies cross-react with sealers |
Add heat and a stain remover such as Tween 20 |
Western Blot Service Advantages
-- For different protein samples, KMD Bioscience adopts different protein concentration determination methods (Lowry method or BCA method)
-- Assurance of the quality of western blot analysis 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 can 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: [email protected] or Tel: +86-022-8216-4980;
FL-4, Building A5, International Enterprise Community, Tianjin, China
Email：[email protected]
Hot Line： +1-315-239-3085
Technical Support：[email protected]
