eIF4E Antibody (JE60-70)

Novus Biologicals | Catalog # NBP3-32302

Recombinant Monoclonal Antibody
Novus Biologicals
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Key Product Details

Species Reactivity

Human, Mouse

Applications

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Flow Cytometry, Immunocytochemistry/ Immunofluorescence

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Rabbit IgG Clone # JE60-70 expressed in HEK293
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Product Specifications

Immunogen

Recombinant protein within Human eIF4E aa 118-217 / 217. (Uniprot: P06730)

Localization

Cytoplasm, P-body, Stress granule, Nucleus.

Clonality

Monoclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

25 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for eIF4E Antibody (JE60-70)

eIF4E Antibody (JE60-70)

Western Blot: eIF4E Antibody (JE60-70) [NBP3-32302]

Western Blot: eIF4E Antibody (JE60-70) [NBP3-32302] - Western blot analysis of eIF4E on different lysates with Rabbit anti-eIF4E antibody (NBP3-32302) at 1/500 dilution.

Lane 1: HL-60 cell lysate
Lane 2: Daudi cell lysate

Lysates/proteins at 10 ug/Lane.

Predicted band size: 25 kDa
Observed band size: 25 kDa

Exposure time: 2 minutes;

15% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (NBP3-32302) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:300,000 dilution was used for 1 hour at room temperature.
eIF4E Antibody (JE60-70)

Immunohistochemistry: eIF4E Antibody (JE60-70) [NBP3-32302]

Immunohistochemistry: eIF4E Antibody (JE60-70) [NBP3-32302] - Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-eIF4E antibody (NBP3-32302) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (NBP3-32302) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
eIF4E Antibody (JE60-70)

Immunocytochemistry/ Immunofluorescence: eIF4E Antibody (JE60-70) [NBP3-32302]

Immunocytochemistry/ Immunofluorescence: eIF4E Antibody (JE60-70) [NBP3-32302] - Immunocytochemistry analysis of SK-Br-3 cells labeling eIF4E with Rabbit anti-eIF4E antibody (NBP3-32302) at 1/50 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-eIF4E antibody (NBP3-32302) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
eIF4E Antibody (JE60-70)

Flow Cytometry: eIF4E Antibody (JE60-70) [NBP3-32302]

Flow Cytometry: eIF4E Antibody (JE60-70) [NBP3-32302] - Flow cytometric analysis of SK-Br-3 cells labeling eIF4E.

Cells were fixed and permeabilized. Then stained with the primary antibody (NBP3-32302, 1ug/ml) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

Applications for eIF4E Antibody (JE60-70)

Application
Recommended Usage

Flow Cytometry

1:500-1:1000

Immunocytochemistry/ Immunofluorescence

1:50

Immunohistochemistry-Paraffin

1:200

Western Blot

1:500

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Advanced Features

  • Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
  • Spillover Popups - Visualize the spectra of individual fluorochromes
  • Antigen Density Selector - Match fluorochrome brightness with antigen density
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Formulation, Preparation, and Storage

Purification

Protein A purified

Formulation

1*TBS (pH7.4), 0.05% BSA and 40% Glycerol

Preservative

0.05% Sodium Azide

Concentration

1 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: eIF4E

eIF-4E is a eukaryotic translation initiation factor involved in directing ribosomes to the cap structure of mRNAs. It exists in two forms: as a free form (25 kDa) and as part of a multiprotein complex eIF-4F (1). eIF-4E appears to be the least abundant of the initiation factors and acts as a rate-limiting step of initiation (2). Since translation is regulated by phosphorylation, eIF-4E phosphorylation at Ser 209 by MAPK signal-integrating kinase 1 (Mnk1) and kinase 2 (Mnk2) may directly regulate the rate of protein synthesis initiation (3). There is also evidence that eIF-4E can function as an oncogene (4-5).

Long Name

Eukaryotic Translation Initiation Factor 4E

Alternate Names

EIF4EL1, EIF4F

Gene Symbol

EIF4E

Additional eIF4E Products

Product Documents for eIF4E Antibody (JE60-70)

Certificate of Analysis

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Product Specific Notices for eIF4E Antibody (JE60-70)

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Related Research Areas

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Protocols

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