Cell Signaling Technology

Product Pathways - Translational Control

Translation Initiation Complex Antibody Sampler Kit #4763

Kit Includes Quantity Applications Reactivity MW (kDa) Isotype
eIF4A (C32B4) Rabbit mAb #2013 40 µl W H M R Mk 48 Rabbit IgG
eIF4A1 Antibody #2490 40 µl W IF-IC H M R Mk 48 Rabbit
Phospho-eIF4B (Ser422) Antibody #3591 40 µl W H M R Mk 80 Rabbit
eIF4B Antibody #3592 40 µl W H M R Mk 80 Rabbit
Phospho-eIF4E (Ser209) Antibody #9741 40 µl W H M R Mk 25 Rabbit
eIF4E (C46H6) Rabbit mAb #2067 40 µl W IP IHC-P H M R Mk 25 Rabbit IgG
Phospho-eIF4G (Ser1108) Antibody #2441 40 µl W IP IF-IC H M R Hm Mk B 220 Rabbit
eIF4G (C45A4) Rabbit mAb #2469 40 µl W IHC-P IF-IC F H M R Mk 220 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl Goat
eIF4H (D85F2) XP® Rabbit mAb #3469 40 µl W IP IF-IC H M R Mk 25, 27 Rabbit IgG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  R=Rat  Hm=Hamster  Mk=Monkey  B=Bovine
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Specificity / Sensitivity

Each antibody in the Translation Initiation Complex Antibody Sampler Kit detects endogenous levels of its target protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using eIF4A (C32B4) Rabbit mAb #2013.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using eIF4E (C46H6) Rabbit mAb #2067.

Western Blotting

Western Blotting

Western blot analysis of extracts from PC12 cells, untreated (lane 1), NGF-treated (10 ng/ml) (lane 2), anisomycin-treated (25 µM) (lane 3), U0126-treated #9903 (10 µM) (lane 4), Rapamycin-treated #9904 (100 nM ) (lane 5) or LY294002-treated #9901 (25 µM) (lane 6), using Phospho-eIF4G (Ser1108) Antibody #2441.


Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using eIF4G (C45A4) Rabbit mAb #2469.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using eIF4A1 Antibody #2490.

Western Blotting

Western Blotting

Western blot analysis of extracts from HepG2, HeLa and NBT-II cells using eIF4H (D85F2) XP® Rabbit mAb #3469.


Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells using Phospho-eIF4B (Ser422) Antibody #3591 (upper) or eIF4B Antibody #3592 (lower). Forty-eight hours following siRNA transfection cells were treated with Rapamycin (50 nM) and U0126 (10 µM) as indicated.

Western Blotting

Western Blotting

Western blot analysis of extracts from NIH/3T3, 293 and HeLa cells using eIF4B Antibody #3592.

Western Blotting

Western Blotting

Western blot analysis of extracts from NIH/3T3 cells, untreated or treated with serum, PD98059 or Dexamethasone, using Phospho-eIF4E (Ser209) Antibody #9741 (upper) or eIF4E Antibody #9742 (lower).


Description

The Translation Initiation Complex Antibody Sampler Kit contains reagents to investigate the initiation of translation within the cell. The kit contains enough primary and secondary antibodies to perform four Western blot experiments per primary antibody.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with a synthetic petide corresponding to residues surrounding Met316 of human eIF4A protein, Gly188 of human eIF4G, and the sequence of human eIF4E and human eIF4H. Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to a sequence around Gly12 of human eIF4A1, residues at the amino terminus of human eIF4B, Ser422 of human eIF4B, Ser209 of human eIF4E, and Ser1108 of human eIF4GI. Polyclonal antibodies are purified by protein A and peptide affinity chromatography.

Background

A variety of factors contribute to the important biological event of translation initiation. The Eukaryotic initiation Factor 4E (eIF4E) complex of translation initiation factors binds to the 5' m7 GTP cap to open up the mRNA secondary structure and allow small ribosome subunit binding (1). eIF4A, an eIF4 complex component that acts as an ATP-dependent RNA helicase, unwinds the secondary structure of the 5' mRNA untranslated region to mediate ribosome binding (2,3). EIF4E binds to the mRNA cap structure to mediate the initiation of translation (4,5). eIF4E interacts with eIF4G, a scaffold protein that promotes assembly of eIF4E and eIF4A into the eIF4F complex (5). eIF4B is thought to assist the eIF4F complex in translation initiation. eIF4H induces the RNA-dependent ATP hydrolysis catalyzed by the initiation factors eIF4A and eIF4B (2,6). eIF4H was further shown to determine the initial rate and extent of eIF4A-mediated mRNA secondary structure unwinding (7).

  1. Rogers, G.W. et al. (2001) J Biol Chem 276, 12598-608.
  2. Rogers, G.W. et al. (1999) J Biol Chem 274, 12236-44.
  3. Svitkin, Y.V. et al. (2001) RNA 7, 382-94.
  4. Sonenberg, N. et al. (1978) Proc Natl Acad Sci U S A 75, 4843-7.
  5. Gingras, A.C. et al. (1999) Annu Rev Biochem 68, 913-63.
  6. Richter-Cook, N.J. et al. (1998) J Biol Chem 273, 7579-87.
  7. Rogers, G.W. et al. (2001) J Biol Chem 276, 30914-22.

Application References

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This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

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