PathScan® Total GFP Sandwich ELISA Kit (#7878) Datasheet with Images Cell Signaling Technology

For Research Use Only. Not for Use in Diagnostic Procedures.

Product Includes	Product #	Quantity	Color	Storage Temp
GFP Rabbit Antibody Coated Microwells	10605	96 tests		+4C
GFP Mouse Detection mAb	14237	1 ea	Green (Lyophilized)	+4C
Anti-mouse IgG, HRP-linked Antibody (ELISA Formulated)	13304	1 ea	Red (Lyophilized)	+4C
Detection Antibody Diluent	13339	11 ml	Green	+4C
HRP Diluent	13515	11 ml	Red	+4C
TMB Substrate	7004	11 ml		+4C
STOP Solution	7002	11 ml		+4C
Sealing Tape	54503	2 ea		+4C
ELISA Wash Buffer (20X)	9801	25 ml		+4C
ELISA Sample Diluent	11083	25 ml	Blue	+4C
Cell Lysis Buffer (10X)	9803	15 ml		-20C

*The microwell plate is supplied as 12 8-well modules - Each module is designed to break apart for 8 tests.

Description

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.

*Antibodies in this kit are custom formulations specific to kit.

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.

Background

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.

Miyawaki, A. et al. (2003) Curr Opin Chem Biol 7, 557-62.

Background References

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

PathScan is a registered trademark of Cell Signaling Technology, Inc.

All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

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