Cell Signaling Technology

Product Pathways - PI3K / Akt Signaling

YB1 (D299) Antibody #4202

Applications Reactivity Sensitivity MW (kDa) Source
W IHC-P IF-IC H M R Mk (X) (B) Endogenous 49 Rabbit

Applications Key:  W=Western Blotting  IHC-P=Immunohistochemistry (Paraffin)  IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  X=Xenopus  B=Bovine
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

YB1 (D299) Antibody detects endogenous levels of total YB1 protein.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to the carboxy terminus of the human YB1 protein. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using YB1 (D299) Antibody.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using YB1 (D299) Antibody in the presence of control peptide (left) or antigen-specific peptide (right).

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells using YB1 (D299) Antibody (green). Actin filaments were labeled using DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).


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).

  1. Matsumoto, K. and Wolffe, A.P. (1998) Trends Cell Biol. 8, 318-23.
  2. Jurchott, K. et al. (2003) J. Biol. Chem. 278, 27988-96.
  3. Mertens, P.R. et al. (1997) J. Biol. Chem. 272, 22905-12.
  4. Uchiumi, T. et al. (1993) Cell Growth Differ. 4, 147-57.
  5. Lasham, A. et al. (2000) Gene 252, 1-13.
  6. Lasham, A. et al. (2003) J. Biol. Chem. 278, 35516-23.
  7. Homer, C. et al. (2005) Oncogene 24, 8314-25.
  8. Raffetseder, U. et al. (2003) J. Biol. Chem. 278, 18241-8.
  9. Chen, C.Y. et al. (2000) Genes Dev. 14, 1236-48.
  10. Sutherland, B.W. et al. (2005) Oncogene 24, 4281-92.

Application References

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For Research Use Only. Not For Use In Diagnostic Procedures.

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