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Anti-EIF3e Antibody Picoband™ (monoclonal, 10F5H6)

Figure 1. Western blot analysis of EIF3E using anti-EIF3E antibody (M00481-1).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human Raji whole cell lysates,
Lane 2: human K562 whole cell lysates,
Lane 3: rat thymus tissue lysates,
Lane 4: mouse thymus tissue lysates,
Lane 5: mouse ANA-1 whole cell lysates,
Lane 6: mouse SP2/0 whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-EIF3E antigen affinity purified monoclonal antibody (Catalog # M00481-1) at 0.5 µg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1001) with Tanon 5200 system. A specific band was detected for EIF3E at approximately 52 kDa. The expected band size for EIF3E is at 52 kDa.
Figure 2. Flow Cytometry analysis of U20S cells using anti-EIF3E antibody (M00481-1).
Overlay histogram showing U20S cells stained with M00481-1 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-EIF3E Antibody (M00481-1, 1 µg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10 µg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1 µg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

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Product Information
Application Applications Legend: E=ELISA WB=Western Blotting IHC=Immunohistochemistry IHC-P=Immunohistochemistry (Paraffin) IP=Immunoprecipitation IF=Immunofluorescence IC=Immunochemistry ICC=Immunocytochemistry FC=Flow Cytometry DB=Dot Blot	WB, FC
Primary Accession	P60228
Host	Mouse
Isotype	Mouse IgG2b
Reactivity	Rat, Human, Mouse
Clonality	Monoclonal
Format	Lyophilized
Description	Anti-EIF3e Antibody Picoband™ (monoclonal, 10F5H6) . Tested in Flow Cytometry, WB applications. This antibody reacts with Human, Mouse, Rat.
Reconstitution	Adding 0.2 ml of distilled water will yield a concentration of 500 µg/ml.

Additional Information
Gene ID	3646
Other Names	Eukaryotic translation initiation factor 3 subunit E {ECO:0000255\|HAMAP-Rule:MF_03004}, eIF3e {ECO:0000255\|HAMAP-Rule:MF_03004}, Eukaryotic translation initiation factor 3 subunit 6 {ECO:0000255\|HAMAP-Rule:MF_03004}, Viral integration site protein INT-6 homolog, eIF-3 p48 {ECO:0000255\|HAMAP-Rule:MF_03004}, EIF3E {ECO:0000255\|HAMAP-Rule:MF_03004}
Calculated MW	52221 Da
Application Details	Western blot, 0.25-0.5 µg/ml, Human, Mouse, Rat Flow Cytometry, 1-3 µg/1x10^6 cells, Human
Source	Eukaryota
Contents	Each vial contains 4 mg Trehalose, 0.9 mg NaCl and 0.2 mg Na2HPO4.
Clone Names	Clone: 10F5H6
Immunogen	E.coli-derived human EIF3e recombinant protein (Position: A160-Q241). Human EIF3e shares 100% amino acid (aa) sequence identity with both mouse and rat EIF3e.
Purification	Immunogen affinity purified.
Storage	At -20°C for one year from date of receipt. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for six months. Avoid repeated freezing and thawing.

For Research Use Only. Not For Use In Diagnostic Procedures.

Protein Information
Name	EIF3E {ECO:0000255\|HAMAP-Rule:MF_03004}
Function	Component of the eukaryotic translation initiation factor 3 (eIF-3) complex, which is required for several steps in the initiation of protein synthesis (PubMed:17581632, PubMed:25849773, PubMed:27462815). The eIF-3 complex associates with the 40S ribosome and facilitates the recruitment of eIF-1, eIF-1A, eIF-2:GTP:methionyl- tRNAi and eIF-5 to form the 43S pre-initiation complex (43S PIC). The eIF-3 complex stimulates mRNA recruitment to the 43S PIC and scanning of the mRNA for AUG recognition. The eIF-3 complex is also required for disassembly and recycling of post-termination ribosomal complexes and subsequently prevents premature joining of the 40S and 60S ribosomal subunits prior to initiation (PubMed:17581632). The eIF-3 complex specifically targets and initiates translation of a subset of mRNAs involved in cell proliferation, including cell cycling, differentiation and apoptosis, and uses different modes of RNA stem-loop binding to exert either translational activation or repression (PubMed:25849773). Required for nonsense-mediated mRNA decay (NMD); may act in conjunction with UPF2 to divert mRNAs from translation to the NMD pathway (PubMed:17468741). May interact with MCM7 and EPAS1 and regulate the proteasome-mediated degradation of these proteins (PubMed:17310990, PubMed:17324924).
Cellular Location	Cytoplasm. Nucleus, PML body.
Tissue Location	Ubiquitously expressed. Expressed at highest levels in appendix, lymph, pancreas, skeletal muscle, spleen and thymus

Research Areas

Application Protocols

Provided below are standard protocols that you may find useful for product applications.

Western Blot Protocol

Blocking Peptides Protocol

Dot Blot Protocol

Immunohistochemistry Protocol

Immunofluorescence Protocol

Immunoprecipitation Protocol

Flow Cytomety Protocol

Cell Culture Protocol

BACKGROUND

Eukaryotic translation initiation factor 3 subunit E is a protein that in humans is encoded by the EIF3E gene. The human homolog of EIF3E is located on chromosome region 8q22-q23. It is composed of 13 exons that span 45 kb of genomic DNA. EIF3E is the component of the eukaryotic translation initiation factor 3 (eIF-3) complex, which is required for several steps in the initiation of protein synthesis ts localization/assembly. The eIF-3 complex associates with the 40S ribosome and facilitates the recruitment of eIF-1, eIF-1A, eIF-2:GTP:methionyl-tRNAi and eIF-5 to form the 43S pre-initiation complex (43S PIC). And the eIF-3 complex stimulates mRNA recruitment to the 43S PIC and scanning of the mRNA for AUG recognition. The eIF-3 complex is also required for disassembly and recycling of post-termination ribosomal complexes and subsequently prevents premature joining of the 40S and 60S ribosomal subunits prior to initiation.

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¥ 3,367.00

Cat# ABO16248

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100 µg

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Anti-EIF3e Antibody Picoband™ (monoclonal, 10F5H6)

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