| Product Includes | Product # | Quantity | Color | Storage Temp |
|---|---|---|---|---|
| YB1 Rabbit mAb Coated Microwells | 87666 | 96 tests |
|
+4C |
| YB1 Mouse Detection mAb | 12627 | 1 ea |
|
+4C |
| Anti-mouse IgG, HRP-linked Antibody (ELISA Formulated) | 13304 | 1 ea |
|
+4C |
| Detection Antibody Diluent | 13339 | 11 ml |
|
+4C |
| HRP Diluent | 13515 | 11 ml |
|
+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 |
|
+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 YB1 Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of YB1 protein. A YB1 rabbit antibody has been coated onto the microwells. After incubation with cell lysates, YB1 protein is captured by the coated antibody. Following extensive washing, a YB1 mouse detection antibody is added to detect the captured YB1 protein. Anti-mouse IgG, HRP-linked antibody is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of the absorbance for the developed color is proportional to the quantity of YB1 protein.
*Antibodies in this kit are custom formulations specific to kit.
Specificity/Sensitivity
Background
The Y-box binding protein 1 (YB1) belongs to a family of evolutionarily conserved, multifunctional Y-box proteins that bind single-stranded DNA and RNA and function as regulators of transcription, RNA metabolism, and protein synthesis (1). YB1 binds to Y-box sequences (TAACC) found in multiple gene promoters and can positively or negatively regulate transcription. YB1 activates genes associated with proliferation and cancer, such as cyclin A, cyclin B1, matrix metalloproteinase-2 (MMP-2), and the multi-drug resistance 1 (MDR1) gene (2-4). YB1 represses genes associated with cell death, including the Fas cell death-associated receptor and the p53 tumor suppressor gene (5-7). It also interacts with the RNA-splicing factor SRp30c and stabilizes interleukin-2 (IL-2) mRNA upon induction of T lymphocytes by IL-2 (8,9). The majority of YB1 protein localizes to the cytoplasm, with a minor pool found in the nucleus; however, nuclear localization appears to be critical for its role in promoting proliferation. Nuclear translocation is cell cycle regulated, with YB1 protein accumulating in the nucleus during G1/S phase (2). In addition, nuclear translocation is induced in response to extracellular stimuli such as hyperthermia and UV irradiation, or treatment of cells with thrombin, interferons, or insulin-like growth factor (IGF-I) (2,10). Treatment of the MCF7 breast cancer cell line with IGF-I results in Akt-mediated phosphorylation of YB1 at Ser102, which is required for nuclear translocation of YB1 and its ability to promote anchorage-independent growth (10). Research studies have shown that YB1 is overexpressed in many malignant tissues, including breast cancer, non-small cell lung carcinoma, ovarian adenocarcinomas, human osteosarcomas, colorectal carcinomas, and malignant melanomas. Investigators have shown that nuclear YB1 expression correlates with high levels of proliferation, drug resistance, and poor tumor prognosis (2,7,10).
- Matsumoto, K. and Wolffe, A.P. (1998) Trends Cell Biol. 8, 318-23.
- Jurchott, K. et al. (2003) J. Biol. Chem. 278, 27988-96.
- Mertens, P.R. et al. (1997) J. Biol. Chem. 272, 22905-12.
- Uchiumi, T. et al. (1993) Cell Growth Differ. 4, 147-57.
- Lasham, A. et al. (2000) Gene 252, 1-13.
- Lasham, A. et al. (2003) J. Biol. Chem. 278, 35516-23.
- Homer, C. et al. (2005) Oncogene 24, 8314-25.
- Raffetseder, U. et al. (2003) J. Biol. Chem. 278, 18241-8.
- Chen, C.Y. et al. (2000) Genes Dev. 14, 1236-48.
- Sutherland, B.W. et al. (2005) Oncogene 24, 4281-92.
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
Limited Uses
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