Background
The green fluorescent protein (GFP) was originally identified as a protein involved in the bioluminescence of the jellyfish Aequorea victoria. GFP cDNA produces a fluorescent product when expressed in prokaryotic cells, without the need for exogenous substrates or cofactors, making GFP a useful tool for monitoring gene expression and protein localization in vivo. Several GFP mutants have been developed, including EGFP, which fluoresce more intensely than the wildtype GFP and have shifted excitation maxima, making them useful for FACS and fluorescence microscopy as well as double-labeling applications. GFP is widely used in expression vectors as a fusion protein tag, allowing expression and monitoring of heterologous proteins fused to GFP.
References
1. Prasher, D.C., et al. 1992. Primary structure of the Aequorea victoria green fluorescent protein. Gene 111: 229-233.
2. Chalfie, M., et al. 1994. Green fluorescent protein as a marker for gene expression. Science 263: 802-805.
Source
This Abmart polyclonal antibody is produced by immunizing animals with a synthetic peptide (KLH-coupled) corresponding to the carboxy terminus of EGFP.
Specificity
GFP Antibody detects GFP and GFP fusion proteins.
Storage
Store at -20°C. Stable for one year from the date of shipment.
Application
-
Western blot analysis of over-expressed GFP protein in COS7 cell lysate, using GFP rabbit polyclonal antibody. The antibody dilutions are 1:1000 (lane1), 1:20-00 (lane2), 1:5000 (lane3), 1:10000 (lane4).
Each lane- was loaded with 10 μg of cell lysate
IP of extracts from COS7 cells transfected with a GFP-tagged protein using GFP rabbit polyclonal antibody (5 μl, lane 3) or using normal rabbit IgG (lane 2) as control, followed by Western blot using Abmart GFP (7G9) Mouse mAb (dilution: 1:5000).
Lane 1 was loaded with 10 μg of COS7 cell lysate over-expressing a GFP-tagged protein.
Reactivity
Isotype
Secondary Antibodies
Use an anti-Rabbit secondary antibody to detect the GFP Antibody.
Recommnded Antibody Dilutions
Western blotting 1:2000
Immunoprecipitation 1:100
|