Figure 1. The relationship between the protein concentration of the lysate from untransfected and GFP transfected HEK 293 cells and the absorbance at 450 nm using the PathScan® Total GFP Sandwich ELISA Antibody Pair #7879 is shown.
|7879S||1 Kit (Reagents for 4 x 96 well plates)||$ 469|
|Product Includes||Volume||Cap Color|
|GFP Rabbit Capture Antibody (100X)||400 µl||Pink|
|GFP Detection Mouse mAb (100X)||400 µl||Blue|
|Anti-mouse IgG, HRP-linked Antibody (1000X)||40 µl||Yellow|
Capture and detection antibodies are stored at 4°C. HRP-linked secondary reagent is stored at -20°C.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
1X Cell Lysis Buffer: 10X Cell Lysis Buffer (#9803): To prepare 10 ml of 1X Cell Lysis Buffer, add 1 ml of 10X Cell Lysis Buffer to 9 ml of dH2O, mix. Buffer can be stored at 4°C for short-term use (1–2 weeks).
Recommended: Add 1 mM phenylmethylsulfonyl fluoride (PMSF) (#8553) immediately before use.
STOP Solution: (#7002)
NOTE: Reagents should be made fresh daily.
posted January 2008
revised Sepetember 2013
Protocol Id: 20
CST's PathScan® Total GFP Sandwich ELISA Antibody Pair is being offered as an economical alternative to our PathScan® Total GFP Sandwich ELISA Kit #7878. This antibody pair is intended for use with cotransfected GFP as a convenient means of monitoring transfection efficiency and may not detect GFP fusion proteins. Capture and detection antibodies (100X stocks) and an HRP-conjugated secondary antibody (1000X stock) are supplied. Sufficient reagents are supplied for 4 x 96 well ELISAs. The GFP rabbit capture antibody is coated onto a 96 well microplate overnight in PBS. After blocking, cell lysate is added followed by a GFP mouse detection antibody and HRP-conjugated, anti-mouse IgG antibody. HRP substrate (TMB) is then added for color development. The magnitude of the absorbance for this developed color is proportional to the quantity of GFP.
For Antibody Pair specificity and sensitivity, please refer to the corresponding PathScan® Sandwich ELISA Kit. Note: This antibody pair detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.
The green-fluorescent protein (GFP) functions as a bioluminescence energy transfer acceptor in the jellyfish Aequorea that maximally absorbs light at 395 nm and has an emission spectrum that peaks at 509 nm (1). Among other applications, GFP is commonly cotransfected alongside a gene of interest as a means of monitoring transfection efficiency.
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