Western blot analysis of GFP was performed by loading various amounts of recombinant GFP per well onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with 5% BSA/TBST for at least 1 hour. The membrane was probed with an HRP-conjugated GFP monoclonal antibody (Product # MA5-15256-HRP) at a dilution of 1:1000 for 1 hour at room temperature on a rocking platform and washed in TBS-0.1% Tween-20. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34080).
|Tested species reactivity||Tag|
|Published species reactivity||Not Applicable|
|Host / Isotype||Mouse / IgG1|
|Immunogen||GFP from the jellyfish Aequorea Victoria N-terminal peptide-KLH conjugated.|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Western Blot (WB)||1:500-1:2000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Western Blot (WB)||See 1 publications below|
MA5-15256-HRP has been successfully used in Western blotting applications with GFP tagged samples.
Epitope tags provide a method to localize gene products in a variety of cell types, study the topology of proteins and protein complexes, identify associated proteins, and characterize newly identified, low abundance or poorly immunogenic proteins when protein specific antibodies are not available. Green Fluorescent Protein (GFP) has quickly become a powerful research tool for assessing gene expression and subcellular protein distribution in fixed or living cells. GFP is excited by and brightly fluoresces when exposed to UV or blue light. This feature makes it ideal as a marker for use in fluorescence microscopy, cytometry, tagging fusion proteins, and assaying transcriptional regulation from gene promoters in vivo. Numerous GFP variants with enhanced and shifted emission spectra (blue, green, and yellow) have been developed through amino acid substitutions at specific residues.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Not Applicable||Not Cited||
c-di-GMP heterogeneity is generated by the chemotaxis machinery to regulate flagellar motility.
MA5-15256-HRP was used in western blot to study the role of the chemotaxis machinery in the partitioning of c-di-GMP concentrations in Pseudomonas aeruginosa
|Kulasekara BR,Kamischke C,Kulasekara HD,Christen M,Wiggins PA,Miller SI||eLife (2:null)||2013|