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 biotinylated GFP monoclonal antibody (Product # MA5-15256-BTIN) at a dilution of 1:1000 for 1 hour at room temperature, followed by Streptavidin-HRP (Product # 21126) at a dilution of 1:20,000 for 30 minutes at room temperature. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34078).
|Tested species reactivity||Tag|
|Host / Isotype||Mouse / IgG1|
|Immunogen||GFP from the jellyfish Aequorea Victoria N-terminal peptide-KLH conjugated.|
|Storage buffer||PBS with proprietary stabilizer|
|Contains||0.02% sodium azide|
|Storage Conditions||4° C, do not freeze|
|Tested Applications||Dilution *|
|ELISA (ELISA)||Assay Dependent|
|Immunoprecipitation (IP)||Assay Dependent|
|Western Blot (WB)||1:500 - 1:1000|
* 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.
MA5-15256-BTIN has been successfully used for Western blot, ELISA, ICC/IF and IP applications.
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.