Purified anti-eIF4B Antibody

Pricing & Availability
Clone
W17131B (See other available formats)
Other Names
Eukaryotic Translation Initiation Factor 4B, EIF-4B, PRO1843
Isotype
Rat IgG2b, κ
Ave. Rating
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Product Citations
publications
W17131B_Purified_eIF4B_Antibody_1_051519.png
Whole cell extracts (15 µg protein) from the indicated cell lines were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with 1.0 µg/mL (1:500 dilution) of Purified anti-eIF4B Antibody, clone W17131B, overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP goat anti-rat IgG Antibody (Cat. No. 405405) at a 1:3000 dilution. Direct-Blot™ HRP anti-GAPDH Antibody (Cat. No. 607904) was used as a loading control at a 1:25000 dilution (lower). Lane M: Molecular Weight marker.
  • W17131B_Purified_eIF4B_Antibody_1_051519.png
    Whole cell extracts (15 µg protein) from the indicated cell lines were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with 1.0 µg/mL (1:500 dilution) of Purified anti-eIF4B Antibody, clone W17131B, overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP goat anti-rat IgG Antibody (Cat. No. 405405) at a 1:3000 dilution. Direct-Blot™ HRP anti-GAPDH Antibody (Cat. No. 607904) was used as a loading control at a 1:25000 dilution (lower). Lane M: Molecular Weight marker.
  • W17131B_Purified_eIF4B_Antibody_2_051519.png
    Whole cell extracts (250 μg total protein) prepared from HeLa cells were immunoprecipitated overnight with 2.5 µg of Purified rat IgG2b, κ isotype Control Antibody (Cat. No. 400602) or Purified anti-eIF4B Antibody, clone W17131B. The resulting IP fractions and whole cell extract input (10%) were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with a 1.0 µg/mL (1:500 dilution) of W17131B. Lane M: Molecular Weight marker.
  • W17131B_Purified_eIF4B_Antibody_3_051519.png
    HeLa cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with ice-cold methanol for 10 minutes, and blocked with 5% FBS for 60 minutes. Cells were then intracellularly stained with a 1:500 dilution (1.0 µg/mL) of either Purified Rat IgG2b, κ isotype Control Antibody (Cat. No. 400602, panel A) or Purified anti-eIF4B Antibody (panel B) overnight at 4°C, followed by incubation with Alexa Fluor® 594 goat anti-rat IgG (Cat. No. 405422) at 2.0 µg/mL. Nuclei were counter-stained with DAPI, and the image was captured with a 60X objective.
Cat # Size Price Quantity Avail. Save
617701 25 µg £57
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617702 100 µg £144
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Description

Eukarytoic initiation factor 4B (eIF4B) is a general translation factor required for the initiation of global protein synthesis. eIF4B facilitates 5’ cap-dependent translation by directly binding to and stimulating the ATPase and RNA-helicase activity of the eIF4F complex. This interaction is regulated by p70 S6 kinase, which phosphorylates eIF4B to promote formation of the eIF4B-eIF4F complex.

Product Details
Technical Data Sheet (pdf)

Product Details

Reactivity
Human, Mouse
Antibody Type
Monoclonal
Host Species
Rat
Immunogen
Partial recombinant human eIF4B protein corresponding to amino acids 1-200
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.
Preparation
The antibody was purified by affinity chromatography.
Concentration
0.5 mg/ml
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C.
Application

WB - Quality tested
ICC, IP - Validated

Recommended Usage

Each lot of this antibody is quality control tested by Western blotting. For Western blotting, the suggested use of this reagent is 0.5 - 1.0 µg per ml. For immunocytochemistry, a concentration range of 1.0 - 5.0 μg/ml is recommended. For immunoprecipitation, the suggested use of this reagent is 2.5 µg per ml. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

When tested for IP, this clone did not produce strong enrichment of eIF4B. Therefore, this clone may not be suitable for using co-IP to detect low abundant or transient protein-protein interactions.

Due to high sequence conservation between rat eIF4B and the immunogen used to generate this clone, we predict this clone will react with the rat eIF4B ortholog.

During PD testing of this clone for ICC, PFA-fixed HeLa cells were permeabilized with either methanol or Triton X-100. Strong staining was observed using both permeabilization methods.

Antigen Details

Structure
eIF4B is a 611 amino acid protein with a predicted molecular weight of 69 kD.
Distribution

Cytosol/Ubiquitously expressed

Function
5' cap-dependent translation initiation
Biology Area
Cell Biology, Protein Synthesis
Antigen References
  1. Grifo JA, et al. 1983. J Biol Chem. 258(9): 5804-10.
  2. Grifo JA, et al. 1984. J. Biol. Chem. 259: 8648.
  3. Holz MK, et al. 2005. Cell. 123:569.
Gene ID
1975 View all products for this Gene ID
UniProt
View information about eIF4B on UniProt.org

Related FAQs

There are no FAQs for this product.
Go To Top Version: 1    Revision Date: 05/15/2019

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