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7940
PathScan® Total eIF4E Sandwich ELISA Kit
ELISA Kits
ELISA Kit

PathScan® Total eIF4E Sandwich ELISA Kit #7940

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PathScan® Total eIF4E Sandwich ELISA Kit: Image 1

Figure 1. Treatment of serum-starved HeLa cells with 20% FBS stimulates phosphorylation of eIF4E at Ser209, detected by the PathScan® Phospho-eIF4E (Ser209) Sandwich ELISA Kit #7938, but does not affect the levels of total eIF4E detected by PathScan® Total eIF4E Sandwich ELISA Kit #7940. HeLa cells (80-90% confluent) were serum-starved over night and then treated with 20% FBS at 37ºC. The absorbance readings at 450 nm are shown in the top figure, while the corresponding western blots using eIF4E Antibody #9742 (left panel) or Phospho-eIF4E (Ser209) Antibody #9741 (right panel) are shown in the bottom figure.

PathScan® Total eIF4E Sandwich ELISA Kit: Image 2

Figure 2. The relationship between the protein concentration of lysates from untreated and 20% FBS-treated HeLa cells and the absorbance at 450 nm using the PathScan® Total eIF4E Sandwich ELISA Kit is shown.

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Supporting Data

REACTIVITY

Application Key:

  • W-Western
  • IP-Immunoprecipitation
  • IHC-Immunohistochemistry
  • ChIP-Chromatin Immunoprecipitation
  • IF-Immunofluorescence
  • F-Flow Cytometry
  • E-P-ELISA-Peptide

Species Cross-Reactivity Key:

  • H-Human
  • M-Mouse
  • R-Rat
  • Hm-Hamster
  • Mk-Monkey
  • 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
  • All-All Species Expected

Product Description

The PathScan® Total eIF4E Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of eIF4E. An eIF4E mouse antibody has been coated onto the microwells. After incubation with cell lysates, eIF4E (phospho and nonphospho) is captured by the coated antibody. Following extensive washing, a eIF4E rabbit detection antibody is added to the captured phospho and nonphospho eIF4E protein. Anti-rabbit IgG, HRP-linked Antibody #7074 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 eIF4E.

Specificity / Sensitivity

CST's PathScan® Total eIF4E Sandwich ELISA Kit detects endogenous levels of total eIF4E protein. As shown in Figure 1, a significant induction of eIF4E phosphorylation at Ser209 can be detected in HeLa cells following treatment with 20% FBS using the Phospho-EIF4E (Ser209) Sandwich ELISA Kit #7938. The levels of total eIF4E remain unchanged as shown by western analysis and by PathScan® Total eIF4E Sandwich ELISA Kit #7940 (Figure 1). This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

Background

Eukaryotic initiation factor 4E (eIF4E) binds to the mRNA cap structure to mediate the initiation of translation (1,2). eIF4E interacts with eIF4G, a scaffold protein that promotes assembly of eIF4E and eIF4A into the eIF4F complex (2). eIF4B is thought to assist the eIF4F complex in translation initiation. Upon activation by mitogenic and/or stress stimuli mediated by Erk and p38 MAPK, Mnk1 phosphorylates eIF4E at Ser209 in vivo (3,4). Two Erk and p38 MAPK phosphorylation sites in mouse Mnk1 (Thr197 and Thr202) are essential for Mnk1 kinase activity (3). The carboxy-terminal region of eIF4G also contains serum-stimulated phosphorylation sites, including Ser1108, Ser1148, and Ser1192 (5). Phosphorylation at these sites is blocked by the PI3 kinase inhibitor LY294002 and by the FRAP/mTOR inhibitor rapamycin.

  1. Sonenberg, N. et al. (1978) Proc Natl Acad Sci U S A 75, 4843-7.
  2. Gingras, A.C. et al. (1999) Annu Rev Biochem 68, 913-63.
  3. Waskiewicz, A. et al. (1999) Mol. Cell. Biol. 19, 1871-80.
  4. Pyronnet, S. et al. (1999) EMBO J. 18, 270-9.
  5. Raught, B. et al. (2000) EMBO J. 19, 434-44.

Pathways & Proteins

Explore pathways + proteins related to this product.

Limited Uses

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Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST's products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST's Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

For Research Use Only. Not For Use In Diagnostic Procedures.
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PathScan is a trademark of Cell Signaling Technology, Inc.