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4763
Translation Initiation Complex Antibody Sampler Kit

Translation Initiation Complex Antibody Sampler Kit #4763

Western Blotting Image 1

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

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Western Blotting Image 2

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

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Western Blotting Image 3

Western blot analysis of extracts from HeLa cells using Phospho-eIF4B (Ser422) Antibody (upper) or eIF4B Antibody #3592 (lower). 48 hours following siRNA transfection, cells were treated with Rapamycin (50 nM) and U0126 (10 µM) as indicated.

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Western Blotting Image 4

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

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Western Blotting Image 5

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

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Western Blotting Image 6

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

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Western Blotting Image 7

Western blot analysis of extracts from 293 cells expressing GST-eIF4GI Ser1192Ala or GST-eIF4GI Ser1108Ala mutant protein, using Phospho-eIF4G (Ser1108) Antibody. (Provided by Brian Raught, McGill University, Montreal, QuŽbec.)

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Western Blotting Image 8

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

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Western Blotting Image 9

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

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Western Blotting Image 10

After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.

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Western Blotting Image 11

Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 (-) or SignalSilence® eIF4E siRNA I #6311 or SignalSilence® eIF4E siRNA II #6554 (+), using eIF4E (C46H6) Rabbit mAb #2067 and α-Tubulin (11H10) Rabbit mAb #2125. The eIF4E (C46H6) Rabbit mAb confirms silencing of eIF4E expression and α-Tubulin (11H10) Rabbit mAb is used to control for loading and specificity of eIF4E siRNA.

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Western Blotting Image 12

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.

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IHC-P (paraffin) Image 13

Immunohistochemical analysis of paraffin-embedded human glioblastoma using eIF4G (C45A4) Rabbit mAb.

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IF-IC Image 14

Confocal immunofluorescent analysis of HeLa cells (left) and MCF-7 cells (right) using eIF4H (D85F2) XP® Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

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IHC-P (paraffin) Image 15

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using eIF4E (C46H6) Rabbit mAb.

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IF-IC Image 16

Confocal immunofluorescent analysis of HeLa cells either rapamycin-treated (left) or serum-treated (right), using Phospho-eIF4G (Ser1108) Antibody (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red). Blue pseudocolor = DRAQ5™ (fluorescent DNA dye).

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IHC-P (paraffin) Image 17

Immunohistochemical analysis of paraffin-embedded human lung carcinoma using eIF4G (C45A4) Rabbit mAb.

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IHC-P (paraffin) Image 18

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using eIF4E (C46H6) Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).

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Flow Cytometry Image 19

Flow cytometric analysis of HeLa cells using eIF4G (C45A4) Rabbit mAb (blue) compared to a nonspecific negative control antibody (red).

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IF-IC Image 20

Confocal immunofluorescent analysis of HeLa cells using eIF4G (C45A4) Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

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Product Includes Quantity Applications Reactivity MW(kDa) Isotype
eIF4A (C32B4) Rabbit mAb 2013 20 µl
  • WB
H M R Mk 48 Rabbit IgG
eIF4A1 Antibody 2490 20 µl
  • WB
H M R Mk 48 Rabbit 
Phospho-eIF4B (Ser422) Antibody 3591 20 µl
  • WB
H M R Mk 80 Rabbit 
eIF4B Antibody 3592 20 µl
  • WB
H M R Mk 80 Rabbit 
Phospho-eIF4E (Ser209) Antibody 9741 20 µl
  • WB
H M R Mk 25 Rabbit 
eIF4E (C46H6) Rabbit mAb 2067 20 µl
  • WB
  • IP
  • IHC
H M R Mk 25 Rabbit IgG
Phospho-eIF4G (Ser1108) Antibody 2441 20 µl
  • WB
  • IP
  • IF
H M R Hm Mk B 220 Rabbit 
eIF4G (C45A4) Rabbit mAb 2469 20 µl
  • WB
  • IHC
  • IF
  • F
H M R Mk 220 Rabbit 
eIF4H (D85F2) XP® Rabbit mAb 3469 20 µl
  • WB
  • IP
  • IF
H M R Mk 25, 27 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

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 two Western blot experiments per primary antibody.

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

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.

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. Sonenberg, N. et al. (1978) Proc. Natl. Acad. Sci. USA 75, 4843-4847.
  3. Rogers, G.W. et al. (1999) J Biol Chem 274, 12236-44.
  4. Gingras, A.C. et al. (1999) Annu. Rev. Biochem. 68, 913-963.
  5. Svitkin, Y.V. et al. (2001) RNA 7, 382-94.
  6. Richter-Cook, N.J. et al. (1998) J Biol Chem 273, 7579-87.
Entrez-Gene Id
1973 , 1974 , 9775 , 1975 , 1977 , 1981 , 7458
Swiss-Prot Acc.
P60842 , Q14240 , P38919 , P23588 , P06730 , Q04637 , Q15056
For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.

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