Revision 2

#7157Store at +4C

 

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Species Cross Reactivity

H M R Mk

UniProt ID:

#P04406

Entrez-Gene Id:

#2597

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Color Storage Temp
GAPDH Rabbit Detection mAb (Biotinylated) 14576 1 ea Green (Lyophilized) +4C
HRP-Linked Streptavidin (ELISA Formulated) 11805 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 Colorless +4C
STOP Solution 7002 11 ml Colorless +4C
Sealing Tape 54503 2 ea +4C
ELISA Wash Buffer (20X) 9801 25 ml Colorless +4C
ELISA Sample Diluent 11083 25 ml Blue +4C
Cell Lysis Buffer (10X) 9803 15 ml Yellowish -20C

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

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.

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.

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.

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.
    XP 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.

    限制使用

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    Revision 2
    #7157

    PathScan® Total GAPDH Sandwich ELISA Kit

    PathScan® Total GAPDH Sandwich ELISA Kit: Image 1 Expand Image
    图 1. 使用 PathScan® Total GAPDH Sandwich ELISA Kit #7157 在多种细胞系中可检测到不同水平的 GAPDH 蛋白。在 450 nm 处的吸光度读数如顶部图所示,使用 GAPDH (D16H11) XP® Rabbit mAb #5174 检测到的相应蛋白质印迹如底部图所示。
    PathScan® Total GAPDH Sandwich ELISA Kit: Image 2 Expand Image
    图 2. HeLa 细胞的裂解物蛋白浓度与使用 PathScan® Total GAPDH Sandwich ELISA Kit #7157 检测的在 450 nm 处的吸光度之间的关系如图所示。HeLa 细胞(85% 融合度)在采集后进行裂解。