This is a knockout-validated antibody summary, based on the two publications "Translational control of nociception via 4E-binding protein 1" [1] and "Translation control during prolonged mTORC1 inhibition mediated by 4E-BP3" [2]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

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Figure 1. Western blot analysis of 4E-BP2 from lysates prepared from DRG and dorsal horn from WT, Eif4ebp1-/- and Eif4ebp2-/- mice. Beta-actin is used as loading control. From [1].

Antibody: 4E-BP2

Catalog number: 2845

Summary: Rabbit polyclonal raised against a synthetic peptide corresponding to residues at the carboxy-terminus of human 4E-BP2. Detects human, mouse, rat, monkey and bovine proteins in western blotting, immunoprecipitation, immunohistochemistry (paraffin), and flow cytometry.

Company: Cell Signaling Technology

Western blot

Lysates prepared from DRG and dorsal horn for 4E-BP1 and 4E-BP2 null mice. Ice-cold homogenization buffer containing (in mM): 20 Tris-HCl, pH 7.4; 150 NaCl; 1 EDTA; 1% Triton, 5 NaF; 1.5 Na3VO4, and protease inhibitor cocktail (complete, EDTA-free, Roche Applied Science).

5% dry milk powder in Tris-buffered saline containing 0.1% Tween-20 (TBS-T) for 1 hr

1:1000 dilution, overnight

HRP-conjugated secondary antibody for 1 h

Enhanced Chemiluminescence reagent (Perkin Elmer), and exposed to an autoradiography film (Denville Scientific Inc.).

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Figure 2. Western blot of cell lysates wild-type and 4E-BP1,2,3 triple knockout MiaPaCa-2 human pancreatic cancer cell lines. Probed with anti-4E-BP2 antibody (row 4)(Cell Signaling Technology, Cat No. 2845). From [2].

Western blot

Lysates of wild-type and knockout MiaPaCa-2 human pancreatic cancer cell lines.

1:1000 dilution.

HRP-conjugated anti-rabbit IgG (Amersham Lifesciences) at 1:1000 dilution.

ECL reagent (GE Healthcare).

If the antibody described in this summary is a polyclonal antibody, since polyclonal antibodies are of limited quantity, please inquire the supplier whether any current polyclonal antibody with the same catalog number is exactly the same as the one described in this summary. Sometimes, different bleeds or different animals are used, usually with a different lot number. In such cases, the result in this summary may not apply to the new antibody with the same catalog number.