Product Pathways - PI3K / Akt Signaling
PathScan® Total YB1 Sandwich ELISA Kit #12543
H M R Hm Mk
|Product Includes||Volume||Solution Color|
|ELISA Sample Diluent||25 ml||Blue|
|YB1 Mouse Detection mAb||1 ea||Green (Lyophilized)|
|Anti-mouse IgG, HRP-linked Antibody (ELISA Formulated)||1 ea||Red (Lyophilized)|
|Detection Antibody Diluent||11 ml||Green|
|HRP Diluent||11 ml||Red|
|STOP Solution||11 ml||Colorless|
|TMB Substrate #7004||11 ml||Colorless|
|ELISA Wash Buffer (20X)||25 ml||Colorless|
|Cell Lysis Buffer (10X) #9803||15 ml||Yellowish|
|YB1 Rabbit mAb Coated Microwells||96 tests|
|Sealing Tape||2 sheets|
Specificity / Sensitivity
PathScan® Total YB1 Sandwich ELISA Kit detects endogenous levels of YB1 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.
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 kit are custom formulations specific to kit.
ELISA - Western correlation
Figure 1. Variable levels of YB1 protein are detected from multiple cell lines using the PathScan® Total YB1 Sandwich ELISA Kit #12543. The absorbance readings at 450 nm are shown in the top figure, while corresponding western blots using YB1 (D2A11) Rabbit mAb #9744 are shown in the bottom figure.
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.
Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!
- 4202 YB1 (D299) Antibody
- 7004 TMB Substrate
- 7076 Anti-mouse IgG, HRP-linked Antibody
- 7249 PathScan® Phospho-YB1 (Ser102) Sandwich ELISA Kit
- 8475 YB1 (D2B12) Rabbit mAb
- 9744 YB1 (D2A11) Rabbit mAb
- 9803 Cell Lysis Buffer (10X)
- 9808 Phosphate Buffered Saline (PBS-20X)
- 9809 Phosphate Buffered Saline with Tween® 20 (PBST-20X)
- 9998 BSA
For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology is a 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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