Phospho-eIF4B (S406) Recombinant Rabbit Monoclonal Antibody [JE63-80]
Catalog# HA722149
Phospho-eIF4B (S406) Recombinant Rabbit Monoclonal Antibody [JE63-80]
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WB
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IF-Cell
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IHC-P
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FC
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Human
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Mouse
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Rat
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unconjugated
Overview
Product Name
Phospho-eIF4B (S406) Recombinant Rabbit Monoclonal Antibody [JE63-80]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic peptide corresponding to residues surrounding Ser406 of human eIF4B protein.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IF-Cell, IHC-P, FC
Target Molecular Weight
Predicted band size: 69 kDa
Positive Control
293T cell lysate, 293T treated with 100nM Calyculin A for 15 minutes cell lysate, HeLa cell lysate, HeLa treated with 100ng/mL Calyculin A for 30 minutes cell lysate, Jurkat cell lysate, Jurkat treated with 100nM Calyculin A for 30 minutes cell lysate, NIH/3T3 cell lysate, NIH/3T3 treated with 100nM Calyculin A for 30 minutes cell lysate, PC-12 cell lysate, PC-12 treated with 100nM Calyculin A for 30 minutes cell lysate, human breast cancer tissue, Jurkat, NIH/3T3, PC-12.
Conjugation
unconjugated
Clone Number
JE63-80
Product Features
Form
Liquid
Concentration
1 mg/mL.(The concentration of this product may be batch-dependent)
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage Buffer
1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:50,000
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IF-Cell
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1:5,000-1:20,000
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IHC-P
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1:1,000
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FC
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1:5,000
Target
Function
Eukaryotic initiation factor 4B (eIF4B) is thought to assist the eIF4F complex in translation initiation. In plants, eIF4B is known to interact with the poly-(A) binding protein, increasing its poly-(A) binding activity. Heat shock and serum starvation cause dephosphorylation of eIF4B at multiple sites with kinetics similar to those of the corresponding inhibition of translation, while phosphorylation of eIF4B following insulin treatment correlates well with an observed increase in translation. Multiple kinases, including p70 S6 kinase, can phosphorylate eIF4B in vitro, and at least one serum-inducible eIF4B phosphorylation site is sensitive to rapamycin and LY294002. Recently, Ser406 was identified as a novel phosphorylation site regulated by mitogens, and the phosphorylation of this site is dependent on MEK and mTOR activity. This phosphorylation is shown to be essential for the translational activity of eIF4B.
Background References
1. Pontheaux F et al. eIF4B mRNA Translation Contributes to Cleavage Dynamics in Early Sea Urchin Embryos. Biology (Basel). 2022 Sep
2. Kapadia BB et al. PARK2 regulates eIF4B-driven lymphomagenesis. Mol Cancer Res. 2022 Feb
Subcellular Location
cytosol, eukaryotic translation initiation factor 4F complex.
Synonyms
2310046H11Rik antibody
AL024095 antibody
C85189 antibody
EIF 4B antibody
eIF-4B antibody
EIF4B antibody
Eukaryotic initiation factor 4B antibody
Eukaryotic translation initiation factor 4B antibody
IF4B_HUMAN antibody
PRO1843 antibody
Images
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☑ Cell treatment (CT)
Western blot analysis of Phospho-eIF4B (S406) on different lysates with Rabbit anti-Phospho-eIF4B (S406) antibody (HA722149) at 1/50,000 dilution.
Lane 1: 293T cell lysate
Lane 2: 293T treated with 100nM Calyculin A for 15 minutes cell lysate
Lane 3: HeLa cell lysate
Lane 4: HeLa treated with 100ng/mL Calyculin A for 30 minutes cell lysate
Lane 5: Jurkat cell lysate
Lane 6: Jurkat treated with 100nM Calyculin A for 30 minutes cell lysate
Lane 7: NIH/3T3 cell lysate
Lane 8: NIH/3T3 treated with 100nM Calyculin A for 30 minutes cell lysate
Lane 9: PC-12 cell lysate
Lane 10: PC-12 treated with 100nM Calyculin A for 30 minutes cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 69 kDa
Observed band size: 80 kDa
Exposure time: 21 seconds; 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 (HA722149) at 1/50,000 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. -
☑ Cell treatment (CT)
Immunohistochemical analysis of paraffin-embedded human breast cancer tissue untreated / treated with λpp with Rabbit anti-Phospho-eIF4B (S406) antibody (HA722149) at 1/1,000 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 (HA722149) at 1/1,000 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. -
☑ Cell treatment (CT)
Immunocytochemistry analysis of Jurkat cells treated with or without λpp labeling Phospho-eIF4B (S406) with Rabbit anti-Phospho-eIF4B (S406) antibody (HA722149) at 1/20,000 dilution.
Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Phospho-eIF4B (S406) antibody (HA722149) at 1/20,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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.
Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Immunocytochemistry analysis of NIH/3T3 cells labeling Phospho-eIF4B (S406) with Rabbit anti-Phospho-eIF4B (S406) antibody (HA722149) at 1/10,000 dilution.
Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Phospho-eIF4B (S406) antibody (HA722149) at 1/10,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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.
Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Immunocytochemistry analysis of PC-12 cells labeling Phospho-eIF4B (S406) with Rabbit anti-Phospho-eIF4B (S406) antibody (HA722149) at 1/10,000 dilution.
Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Phospho-eIF4B (S406) antibody (HA722149) at 1/10,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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.
Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Flow cytometric analysis of Jurkat cells labeling Phospho-eIF4B (S406).
Cells were fixed and permeabilized. Then stained with the primary antibody (HA722149, 1/5,000) (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 (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). -
Flow cytometric analysis of NIH/3T3 cells labeling Phospho-eIF4B (S406).
Cells were fixed and permeabilized. Then stained with the primary antibody (HA722149, 1/5,000) (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™ 594 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1122) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). -
Flow cytometric analysis of PC-12 cells labeling Phospho-eIF4B (S406).
Cells were fixed and permeabilized. Then stained with the primary antibody (HA722149, 1/5,000) (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™ 594 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1122) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
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