Overview

  • Product name

    Human eIF4E ELISA Kit
    See all eIF4E kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    MCF7 extract 8 3.4%
    Inter-assay
    Sample n Mean SD CV%
    MCF7 extract 3 10%
  • Sample type

    Cell culture extracts, Tissue Extracts
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    34.69 pg/ml
  • Range

    109.4 pg/ml - 7000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Tissue Extracts 108 106% - 111%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Human
  • Product overview

    eIF4E in vitro SimpleStep ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of eIF4E protein in human cell and tissue extracts.


    The SimpleStep ELISA employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.

  • Notes

    Eukaryotic translation initiation factor 4E (eIF4E) plays a role in the initial stages of translation by recognizing the 7-methyl-guanosine cap on the 5’ end of mRNA. By unwinding mRNA secondary structure, it facilitates ribosomal recruitment. eIF4E is also a part of a complex containing CYFIP1 and FMR1 that can repress translation.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Pre-coated microplate (12 x 8 well strips)

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Wash Buffer PT (ab206977) 1 x 20ml
    50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml
    5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml
    Antibody Diluent 5BR 1 x 6ml
    10X Human eIF4E Capture Antibody 1 x 600µl
    20X Human eIF4E Detector Antibody 1 x 330µl
    Human eIF4E Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS 1 x 12ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • Research areas

  • Function

    Its translation stimulation activity is repressed by binding to the complex CYFIP1-FMR1 (By similarity). Recognizes and binds the 7-methylguanosine-containing mRNA cap during an early step in the initiation of protein synthesis and facilitates ribosome binding by inducing the unwinding of the mRNAs secondary structures. Component of the CYFIP1-EIF4E-FMR1 complex which binds to the mRNA cap and mediates translational repression. In the CYFIP1-EIF4E-FMR1 complex this subunit mediates the binding to the mRNA cap.
  • Sequence similarities

    Belongs to the eukaryotic initiation factor 4E family.
  • Post-translational
    modifications

    Phosphorylation increases the ability of the protein to bind to mRNA caps and to form the eIF4F complex.
  • Information by UniProt
  • Alternative names

    • AUTS19
    • CBP
    • eIF 4E
    • EIF 4F
    • eIF 4F 25 kDa subunit
    • eIF-4E
    • eIF-4F 25 kDa subunit
    • eIF4E
    • EIF4E1
    • EIF4EL1
    • EIF4F
    • Eukaryotic translation initiation factor 4 E
    • Eukaryotic translation initiation factor 4E
    • Eukaryotic translation initiation factor 4E like 1
    • IF4E_HUMAN
    • Messanger RNA Cap Binding Protein eIF 4E
    • MGC111573
    • mRNA cap binding protein
    • mRNA cap-binding protein
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab214564 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Sandwich ELISA Use at an assay dependent concentration.

Images

  • SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

     

  • Background-subtracted data values (mean +/- SD) are graphed.

  • The concentrations of eIF4E were measured in duplicate and interpolated from the eIF4E standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean eIF4E concentration was determined to be 4,667 pg/mL in MCF7, 2,899 pg/mL in HL‑60, and 3,021 pg/mL in placenta.

Protocols

References

ab214564 has not yet been referenced specifically in any publications.

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