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Phospho-eIF4EBP1 (T70) Recombinant Rabbit Monoclonal Antibody [PSH09-91]

Usd: 385

Specification

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Catalog# HA723222

Phospho-eIF4EBP1 (T70) Recombinant Rabbit Monoclonal Antibody [PSH09-91]

Application

  • WB

  • IHC-P

  • IP

Reactivity

  • Human

  • Mouse

BSA and Azide free
Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

Phospho-eIF4EBP1 (T70) Recombinant Rabbit Monoclonal Antibody [PSH09-91]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic phospho-peptide corresponding to residues surrounding Thr70 of human eIF4EBP1.

Species Reactivity

Human, Mouse

Validated Applications

WB, IHC-P, IP

Molecular Weight

Predicted band size: 13 kDa

Positive Control

HEK-293 cell lysate, HEK-293 treated with 20% FBS for 3 hours cell lysate, mouse liver tissue, mouse pancreas tissue.

Conjugation

unconjugated

Clone Number

PSH09-91

Reactivity Data

WB IHC-P IP
Human
Tested Tested
Not Recommended
Tested Tested
Mouse
Not Recommended
Tested Tested
Rat
Not Recommended
Not Recommended

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:2,000

  • IHC-P

  • 1:200

  • IP

  • 1-2μg/sample

Target

Function

This gene encodes one member of a family of translation repressor proteins. The protein directly interacts with eukaryotic translation initiation factor 4E (eIF4E), which is a limiting component of the multisubunit complex that recruits 40S ribosomal subunits to the 5' end of mRNAs. Interaction of this protein with eIF4E inhibits complex assembly and represses translation. This protein is phosphorylated in response to various signals including UV irradiation and insulin signaling, resulting in its dissociation from eIF4E and activation of cap-dependent mRNA translation.

Background References

1. Voeltzke K et al. EIF4EBP1 is transcriptionally upregulated by MYCN and associates with poor prognosis in neuroblastoma. Cell Death Discov. 2022 Apr

2. Nelson ED et al. High EIF4EBP1 expression reflects mTOR pathway activity and cancer cell proliferation and is a biomarker for poor breast cancer prognosis. Am J Cancer Res. 2024 Jan

Subcellular Location

Cytoplasm, cytosol.

UNIPROT

SwissProt: Q13541 Human

SwissProt: Q60876 Mouse

Synonyms

4E-BP1 antibody

4EBP1 antibody

4EBP1_HUMAN antibody

BP 1 antibody

eIF4E binding protein 1 antibody

eIF4E-binding protein 1 antibody

Eif4ebp1 antibody

Eukaryotic translation initiation factor 4E-binding protein 1 antibody

PHAS-I antibody

PHASI antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of Phospho-eIF4EBP1 (T70) on different lysates with Rabbit anti-Phospho-eIF4EBP1 (T70) antibody (<a href="/products/HA723222" style="font-weight: bold;text-decoration: underline;">HA723222</a>) at 1/2,000 dilution.<br /><br />Lane 1: HEK-293 cell lysate<br />Lane 2: HEK-293 treated with 20% FBS for 3 hours cell lysate<br />Lane 3: HEK-293 treated with 20% FBS for 3 hours cell lysate, then the membrane treated with λpp for 1 hour<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 13 kDa<br />Observed band size: 17/18 kDa<br /><br />Exposure time: 3 minutes; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA723222" style="font-weight: bold;text-decoration: underline;">HA723222</a>) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Cell treatment (CT)

    Western blot analysis of Phospho-eIF4EBP1 (T70) on different lysates with Rabbit anti-Phospho-eIF4EBP1 (T70) antibody (HA723222) at 1/2,000 dilution.

    Lane 1: HEK-293 cell lysate
    Lane 2: HEK-293 treated with 20% FBS for 3 hours cell lysate
    Lane 3: HEK-293 treated with 20% FBS for 3 hours cell lysate, then the membrane treated with λpp for 1 hour

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 13 kDa
    Observed band size: 17/18 kDa

    Exposure time: 3 minutes; ECL: K1801;

    4-20% 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 (HA723222) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-Phospho-eIF4EBP1 (T70) antibody (<a href="/products/HA723222" style="font-weight: bold;text-decoration: underline;">HA723222</a>) at 1/200 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA723222" style="font-weight: bold;text-decoration: underline;">HA723222</a>) 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.

    Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-Phospho-eIF4EBP1 (T70) antibody (HA723222) 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 (HA723222) 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.

  • Immunohistochemical analysis of paraffin-embedded mouse pancreas tissue with Rabbit anti-Phospho-eIF4EBP1 (T70) antibody (<a href="/products/HA723222" style="font-weight: bold;text-decoration: underline;">HA723222</a>) at 1/200 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA723222" style="font-weight: bold;text-decoration: underline;">HA723222</a>) 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.

    Immunohistochemical analysis of paraffin-embedded mouse pancreas tissue with Rabbit anti-Phospho-eIF4EBP1 (T70) antibody (HA723222) 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 (HA723222) 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.

  • Phospho-eIF4EBP1 (T70) was immunoprecipitated from 0.2 mg HEK-293 cell lysate with <a href="/products/HA723222" style="font-weight: bold;text-decoration: underline;">HA723222</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/HA723222" style="font-weight: bold;text-decoration: underline;">HA723222</a> at 1/2,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.<br /><br />Lane 1: HEK-293 cell lysate (input)<br />Lane 2: <a href="/products/HA723222" style="font-weight: bold;text-decoration: underline;">HA723222</a> IP in HEK-293 cell lysate<br />Lane 3: Rabbit IgG instead of <a href="/products/HA723222" style="font-weight: bold;text-decoration: underline;">HA723222</a> in HEK-293 cell lysate<br /><br />Blocking/Dilution buffer: 5% NFDM/TBST<br />Exposure time: 3 minutes; ECL: <a href="/products/K1802" style="font-weight: bold;text-decoration: underline;">K1802</a>

    Phospho-eIF4EBP1 (T70) was immunoprecipitated from 0.2 mg HEK-293 cell lysate with HA723222 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA723222 at 1/2,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.

    Lane 1: HEK-293 cell lysate (input)
    Lane 2: HA723222 IP in HEK-293 cell lysate
    Lane 3: Rabbit IgG instead of HA723222 in HEK-293 cell lysate

    Blocking/Dilution buffer: 5% NFDM/TBST
    Exposure time: 3 minutes; ECL: K1802

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Citation