Product Pathways - Metabolism
PathScan® Total GAPDH Sandwich ELISA Kit #7157
H M R Mk
|Product Includes||Volume||Solution Color|
|ELISA Sample Diluent||25 ml||Blue|
|HRP-Linked Streptavidin (ELISA Formulated)||1 ea||Red (Lyophilized)|
|Detection Antibody Diluent||11 ml||Green|
|HRP Diluent||11 ml||Red|
|GAPDH Rabbit Detection mAb (Biotinylated)||1 ea||Green (Lyophilized)|
|GAPDH Rabbit mAb Coated Microwells||96 tests|
|STOP Solution #7002||11 ml||Colorless|
|TMB Substrate #7004||11 ml||Colorless|
|ELISA Wash Buffer (20X)||25 ml||Colorless|
|Cell Lysis Buffer (10X) #9803||15 ml||Yellowish|
|Sealing Tape||2 sheets|
Specificity / Sensitivity
PathScan® Total GAPDH Sandwich ELISA Kit detects endogenous levels of GAPDH protein as shown in Figure 1. The kit sensitivity is shown in Figure 2. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.
PathScan® Total GAPDH Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of GAPDH. A GAPDH Rabbit mAb has been coated onto the microwells. After incubation with cell lysates, GAPDH is captured by the coated antibody. Following extensive washing, a biotinylated GAPDH Rabbit Detection mAb is added to detect the captured phospho and nonphospho GAPDH protein. A HRP-linked streptavidin antibody is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of the absorbance for this developed color is proportional to the quantity of total GAPDH.
Antibodies in kit are custom formulations specific to kit.
ELISA - Western correlation
Figure 1. GAPDH protein is detected at varying levels from multiple cell lines using the PathScan® Total GAPDH Sandwich ELISA Kit #7157. The absorbance readings at 450 nm are shown in the top figure, while the corresponding western blots using GAPDH (D16H11) XP® Rabbit mAb #5174 are shown in the bottom figure.
Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) catalyzes the phosphorylation of glyceraldehyde-3-phosphate during glycolysis. Though differentially expressed from tissue to tissue (1), GAPDH is thought to be a constitutively expressed housekeeping protein. For this reason, GAPDH mRNA and protein levels are often measured as controls in experiments quantifying specific changes in expression of other targets. Recent work has elucidated roles for GAPDH in apoptosis (2), gene expression (3), and nuclear transport (4). GAPDH may also play a role in neurodegenerative pathologies such as Huntington and Alzheimer's diseases (4,5).
- Barber, R.D. et al. (2005) Physiol. Genomics 21, 389-95.
- Hara, M.R. and Snyder, S.H. (2006) Cell Mol. Neurobiol. 26, 527-38.
- Zheng, L. et al. (2003) Cell 114, 255-66.
- Bae, B.I. et al. (2006) Proc. Natl. Acad. Sci. USA 103, 3405-9.
- Wang, Q. et al. (2005) FASEB J. 19, 869-71.
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- 2118 GAPDH (14C10) Rabbit mAb
- 5174 GAPDH (D16H11) XP® Rabbit mAb
- 7002 STOP Solution
- 7004 TMB Substrate
- 9803 Cell Lysis Buffer (10X)
- 9809 Phosphate Buffered Saline with Tween® 20 (PBST-20X)
- 9998 BSA
For Research Use Only. Not For Use In Diagnostic Procedures.
U.S. Patent No. 5,675,063.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
PathScan is a trademark of Cell Signaling Technology, Inc.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.
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