Cell Signaling Technology

Product Pathways - Metabolism

PathScan® Total GAPDH Sandwich ELISA Kit #7157

elisa   GAPDH   sandwich elisa   screen  

REACTIVITY
H M R Mk

No. Size Price
7157C 1 Kit ( 96 assays ) ¥6,338.00 现货查询 购买询价
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.

Description

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

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.

Sensitivity

Sensitivity

Figure 2. The relationship between protein concentration of lysates from HeLa cells and the absorbance at 450 nm as detected by the PathScan® Total GAPDH Sandwich ELISA Kit #7157 is shown. HeLa cells (85% confluence) were harvested and then lysed.

Background

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).

  1. Barber, R.D. et al. (2005) Physiol. Genomics 21, 389-95.
  2. Hara, M.R. and Snyder, S.H. (2006) Cell Mol. Neurobiol. 26, 527-38.
  3. Zheng, L. et al. (2003) Cell 114, 255-66.
  4. Bae, B.I. et al. (2006) Proc. Natl. Acad. Sci. USA 103, 3405-9.
  5. Wang, Q. et al. (2005) FASEB J. 19, 869-71.

Application References

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Protocols

Companion Products


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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