EIF4E Monoclonal antibody

• Featured Product
• KD/KO Validated

EIF4E Monoclonal Antibody for WB, IHC, ELISA, IF/ICC, FC (Intra)

Host / Isotype

Mouse / IgG1

Reactivity

Human, mouse, rat and More (1)

Applications

WB, IHC, ELISA, IF/ICC, FC (Intra)

Conjugate

Unconjugated

Publications

4

CloneNo.

3C6B9

Cat no : 66655-1-Ig

Synonyms

eIF-4F 25 kDa subunit, EIF4F, EIF4EL1, eIF-4E, eIF 4E

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Validation Data Gallery

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  WB analysis using 66655-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 66655-1-Ig (EIF4E antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours. The membrane was stripped and reblotted with HRP-conjugated GAPDH Monoclonal antibody (HRP-60004) as loading control.

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  WB analysis of HEK-293 using 66655-1-Ig

  WB result of EIF4E antibody (66655-1-Ig; 1:10000; incubated at room temperature for 1.5 hours) with sh-Control and sh-EIF4E transfected HEK-293 cells.

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  WB analysis using 66655-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 66655-1-Ig (EIF4E antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours. The membrane was stripped and reblotted with HRP-conjugated GAPDH Monoclonal antibody (HRP-60004) as loading control.

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  WB analysis of HeLa using 66655-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 66655-1-Ig (EIF4E antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

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  WB analysis of U2OS using 66655-1-Ig

  U2OS cells were subjected to SDS PAGE followed by western blot with 66655-1-Ig (EIF4E antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours.

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  IHC staining of human breast cancer using 66655-1-Ig

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 66655-1-Ig (EIF4E antibody) at dilution of 1:5000 (under 40x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human breast cancer using 66655-1-Ig

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 66655-1-Ig (EIF4E antibody) at dilution of 1:5000 (under 10x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of HepG2 using 66655-1-Ig

  Immunofluorescent analysis of (-20°C Ethanol) fixed HepG2 cells using 66655-1-Ig (EIF4E antibody) at dilution of 1:100 and Alexa Fluor 488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L).

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  FC experiment of HepG2 using 66655-1-Ig

  1X10^6 HepG2 cells were intracellularly stained with 0.2 ug Anti-Human EIF4E (66655-1-Ig, Clone:3C6B9) and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L) at dilution 1:1000 (red), or 0.2 ug Control Antibody. Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

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Tested Applications

• Positive WB detected in: Jurkat cells, U2OS cells, LNCaP cells, HeLa cells, HEK-293 cells, HSC-T6 cells, NIH/3T3 cells, RAW 264.7 cells, MCF-7 cells
• Positive IHC detected in: human breast cancer tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF/ICC detected in: HepG2 cells
• Positive FC (Intra) detected in: HepG2 cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunohistochemistry (IHC): IHC : 1:2500-1:10000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:50-1:500
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.20 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• KD/KO: See 1 publications below
• WB: See 3 publications below
• IHC: See 1 publications below
• IF: See 1 publications below

Product Information

66655-1-Ig targets EIF4E in WB, IHC, ELISA, IF/ICC, FC (Intra) applications and shows reactivity with Human, mouse, rat samples.

• Tested Reactivity: Human, mouse, rat
• Cited Reactivity: human, mouse
• Host / Isotype: Mouse / IgG1
• Class: Monoclonal
• Type: Antibody
• Immunogen: EIF4E fusion protein Ag27191
• Full Name: eukaryotic translation initiation factor 4E
• Calculated Molecular Weight: 29 kDa
• Observed Molecular Weight: 26-29 kDa
• GenBank Accession Number: BC012611
• Gene Symbol: EIF4E
• Gene ID (NCBI): 1977
• RRID: AB_2882012
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

Eukaryotic translation initiation factor 4E, also known as eIF4E, is a protein that in humans is encoded by the EIF4E gene. eIF4E is the mRNA cap-binding protein, known as a general initiation factor allowing for mRNA-ribosome interaction and cap-dependent translation in eukaryotic cells. eIF4E is a polypeptide that exists as both a free form and as part of the eIF4F pre-initiation complex. Regulation of eIF4E may be achieved via three distinct mechanisms: transcription, phosphorylation, and inhibitory proteins.

Publications

Species	Application	Title
human	WB	J Immunother Cancer; Genome-scale CRISPR-Cas9 screen reveals novel regulators of B7-H3 in tumor cells.; Authors - Shasha Zhao
mouse	WB,IHC	Neurosci Bull; miR-34b-3p Inhibition of eIF4E Causes Post-stroke Depression in Adult Mice.; Authors - Xiao Ke ; KD Validated
human	IF	J Cell Biol; DDX6 modulates P-body and stress granule assembly, composition, and docking; Authors - Nina Ripin
human	WB	World J Gastroenterol; Thymoquinone affects hypoxia-inducible factor-1α expression in pancreatic cancer cells via HSP90 and PI3K/AKT/mTOR pathways; Authors - Zhan-Xue Zhao

Related Products

Flow Cytometry Perm Buffer (10X)

PF00011-C