Product Pathways - PathScan ELISA
PathScan® Total GFP Sandwich ELISA Kit #7878
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|Kit Includes||Volume||Solution Color|
|GFP Ab Coated Microwells||96 tests|
|GFP Detection Ab||11 ml||Green|
|Anti-mouse IgG, HRP-linked Antibody||11 ml||Red|
|Sample Diluent||25 ml||Blue|
|TMB Substrate #7004||11 ml||Colorless|
|STOP Solution #7002||11 ml||Colorless|
|Sealing Tape||2 sheets|
|ELISA Wash Buffer (20X)||25 ml||Colorless|
|Cell Lysis Buffer (10X) #9803||15 ml||Yellowish|
Note: 12 8-well modules –Each module is designed to break apart for 8 tests.
Note: Kit should be stored at 4°C with the exception of Cell Lysis Buffer, which is stored at –20°C (packaged separately).
Reactivity Key: All=All species expected
The PathScan® Total GFP Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects exogenously expressed GFP. This kit is intended for use with cotransfected GFP as a convenient means of monitoring transfection efficiency. A GFP rabbit antibody has been coated onto the microwells. After incubation with cell lysates, GFP is captured by the coated antibody. Following extensive washing, a GFP mouse detection antibody is added to detect the captured GFP. Anti-mouse IgG, HRP-linked antibody is then used to recognize the bound detection antibody. The HRP substrate TMB is added to develop color. The magnitude of the absorbance for this developed color is proportional to the quantity of GFP.
Specificity / Sensitivity
CST's PathScan® Total GFP Sandwich ELISA Kit #7878 detects GFP when exogenously expressed in cells. As shown in Figure 1, this kit readily detects GFP in transfected, but not in untransfected HEK293 cell lysate. This kit was developed for use with transfected full-length GFP and may not detect GFP fusion proteins. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.
ELISA - Western correlation
Figure 1. Transfected GFP is detected by PathScan® Total GFP Sandwich ELISA Kit #7878. Lysates were prepared from control and GFP-transfected HEK293 cells and analyzed for GFP by PathScan® Total GFP Sandwich ELISA Kit #7878 or by Western blotting. Absorbance readings at 450 nm are shown in the top figure while the corresponding Western blot using GFP Antibody #2555 is shown in the bottom figure.
The green-fluorescent protein (GFP) functions as a bioluminescence energy transfer acceptor in the jellyfish Aequorea. GFP 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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- 2555 GFP Antibody
- 2956 GFP (D5.1) XP® Rabbit mAb
- 7076 Anti-mouse IgG, HRP-linked Antibody
- 9803 Cell Lysis Buffer (10X)
- 7004 TMB Substrate
- 7002 STOP Solution
- 9808 Phosphate Buffered Saline (PBS-20X)
- 9809 Phosphate Buffered Saline with Tween 20 (PBST-20X)
For Research Use Only. Not For Use In Diagnostic Procedures.