Western Blotting Quantification Service

Western blotting quantification is a protein expression detection method that combines western blotting technology with high-sensitivity quantitative analysis. After proteins are separated by electrophoresis based on their size, they are transferred to a nitrocellulose or PVDF membrane and detected through specific antibody-antigen interactions. Western blotting quantification not only enables qualitative analysis of proteins but also provides reliable protein expression data through precise quantification, addressing the uncertainties associated with traditional western blotting analysis. By utilizing advanced imaging technologies and quantitative analysis software, researchers can obtain accurate protein expression data, thereby advancing studies in protein function, disease mechanisms, and drug development.

 

MtoZ Biolabs offers an efficient and precise western blotting quantification service that leverages advanced technological platforms and a professional team to provide reliable protein expression data, supporting in-depth research in molecular biology, biochemistry, and disease studies.

 

Analysis Workflow

To ensure accurate and reliable results, MtoZ Biolabs' western blotting quantification service follows a meticulous workflow:

1. Sample Preparation: Extract and quantify proteins from cells, tissues, or biological samples for subsequent gel loading.

2. Protein Separation: Use SDS-PAGE to separate target proteins based on molecular weight, forming distinct bands.

3. Transfer and Blocking: Transfer proteins onto PVDF or nitrocellulose membranes and block nonspecific binding sites.

4. Antibody Detection: Add primary antibodies for target binding, followed by secondary antibodies to amplify the signal.

5. Signal Detection: Record and analyze signals using chemiluminescence (ECL) or fluorescence imaging systems.

6. Data Analysis and Reporting: Provide a detailed report with experimental procedures, result images, and data interpretation.

 



 

Applications

Western blotting quantification service by MtoZ Biolabs is an indispensable tool for advancing research in molecular biology and biomedical science. This service enables precise analysis of protein expression, supporting critical applications such as investigating signaling pathways, studying gene expression regulation, exploring protein functions, conducting clinical research, and understanding pathophysiological mechanisms. With MtoZ Biolabs' expertise and cutting-edge technology, researchers gain high-quality, reliable data to drive discoveries in these essential areas of study.

 

Case Study

Case 1:
Western blotting plays a critical role in detecting protein modifications. In one study, enhanced chemiluminescence (ECL) technology was combined with anti-phosphotyrosine antibodies to semi-quantitatively analyze phosphorylated proteins. The researchers prepared and validated a phosphotyrosine protein standard as a reference. Protein tyrosine phosphorylation is a key process in activating receptor tyrosine kinases (RTKs), which are central drivers in the development of certain cancers. This case highlights the high sensitivity and accuracy of western blotting in analyzing phosphorylated proteins, emphasizing its importance in signal pathway research, cancer mechanism studies, and protein modification analysis.



Kendrick, N. et al. PLOS ONE. 2020.

 

Case 2:
Western blotting was used to analyze skeletal muscle biopsy samples from healthy controls and patients with Duchenne muscular dystrophy (DMD), successfully quantifying the expression levels of mini-dystrophin. The results showed a significant increase in protein expression after treatment, with differences in the expression pattern compared to healthy individuals. This study demonstrated that mini-dystrophin could serve as an evaluation marker for gene therapy efficacy. It also highlighted the high sensitivity and reliability of western blotting in quantifying low-abundance proteins and analyzing clinical samples, proving its value in gene therapy research.

 

Soderstrom, C. I. et al. AAPS J. 2022.

 

Case 3:
Western blotting was applied to detect O-GlcNAc-modified proteins that had been chemically labeled using enzymatic mass tagging (CMT) and strain-promoted azide-alkyne cycloaddition (SPAAC) reactions. Proteins were separated by SDS-PAGE and quantitatively analyzed to measure the stoichiometric ratios of O-GlcNAc modifications. This method validated the labeling efficiency and quantitatively reflected dynamic changes in modification levels. The results demonstrated the technique’s critical role in exploring the functions of O-GlcNAc modifications, evaluating their dynamics, and understanding their biological significance. 

 



Darabedian, .N. et al. Biochemistry.

 

Case 4:
Western blotting was used to quantify the expression levels of P-glycoprotein (P-gp) in gastrointestinal tissues from humans and rodents. The results showed the highest expression levels in the small intestine, particularly in the jejunum and ileum, and relatively lower expression in the stomach and colon. Furthermore, significant differences were observed in P-gp expression levels across genders and species. As a complementary method to LC-MS/MS, western blotting provided reliable quantitative data, confirming the regional specificity of P-gp expression and revealing its variability between genders and species. This study supports the application of western blotting in drug metabolism and transport research.

 

 

Mai, Y. et al. Mol. Pharm. 2021.

 

Why Choose MtoZ Biolabs?

1. Advanced Analysis Platform: MtoZ Biolabs established an advanced western blotting quantification analysis platform, guaranteeing reliable, fast, and highly accurate analysis service.

2. One-Time-Charge: Our pricing is transparent, no hidden fees or additional costs.

3. High-Data-Quality: Deep data coverage with strict data quality control. AI-powered bioinformatics platform integrate all western blotting quantification analysis data providing clients with a comprehensive data report.

 

What Could be Included in the Report?

1. Comprehensive Experimental Details

2. Materials, Instruments, and Methods

3. Western Blot Imaging and Quality Control Assessment

4. Data Analysis and Quantification

5. Bioinformatics and Statistical Analysis

6. Raw Data Files

 

For a free project evaluation, contact us today! MtoZ Biolabs technical specialists are ready to assist with your western blotting quantification service needs.