Western Blot Analysis Services

Protein Extraction Services:

• Extraction and purification of protein from tissues provided by customers or Zyagen
• Extraction and purification of protein from primary cell or cell lines provided by customers
• Protein is extracted in any lysis buffer suggested by customers
• To prevent protein degradation, protein lysis buffer is supplemented with a cocktail of 7 protease inhibitors.
• Quantification of protein using highly specific and sensitive protein assays.
• Quality of protein is tested by denatured SDS gel electrophoresis
• Extracted protein is suitable for immediate use in reporter assays (e.g., luciferase, beta-galactosidase, chloramphenicol acetyl transferase), protein kinase assays (e.g., PKA, PKC, tyrosine kinase), immunoassays (e.g., Western blots, ELISAs, RIAs) and/or protein purification procedures, immunoprecipitation,
• SDS-PAGE, isoelectric focusing gels and SDS-capillary electrophoresis, enzymatic activity analysis, gel shifting assay, and protein-protein interaction.
• Protein is provided in protein lysis buffer or denatured SDS sample buffer at any concentration suggested by customers
• Protein is shipped on dry ice

Construction of Premade Western blotting Services:

• Harvesting fresh tissues/cultured cells provided by Zyagen or customers.
• Homogenization of tissues/cells in protein lysis buffer supplemented with a cocktail of 7 mammalian protease inhibitors.
• Quantitation of  total protein using sensitive, specific and accurate protein assay.
• Testing the purity and integrity of extracted proteins by gel electrophoresis.
• Fractionation of protein (75 microgram) through large SDS-denatured polyacrylamide gel electrophoresis.
• Transfer protein onto PVDF membranes.
• Staining blots with reversible staining to check the sharpness of protein bands and efficiency of transfer.
• Shipping blots at room temperature in sealed bags

Western Blot Analysis Services:

• Extraction of total proteins from tissues/cells provided by Zyagen or customers
• Constructions of Western blots using total proteins
• Immunostaining blots with antibodies provided by customers using ultrasensative chemiluminescent ECL detection
• Stripping antibodies from blots
• Reimmunostain blots with any house-keeping gene antibodies
• Scanning protein bands on X-ray films
• Quantitative analysis of immunostaining data
• Sending high quality digital images of X-ray films on Microsoft PowerPoint