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Product Info Summary
| SKU: | A00481-1 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Rabbit |
| Application: | IP, IHC, WB |
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Product info
Product Name
Anti-EIF3e Antibody Picoband®
SKU/Catalog Number
A00481-1
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-EIF3e Antibody Picoband® catalog # A00481-1. Tested in IP, IHC, WB applications. This antibody reacts with Human, Mouse, Rat. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
Storage & Handling
Store 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 freeze-thaw cycles.
Cite This Product
Anti-EIF3e Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00481-1)
Host
Rabbit
Contents
Each vial contains 4 mg Trehalose, 0.9 mg NaCl and 0.2 mg Na2HPO4.
Clonality
Polyclonal
Isotype
Rabbit IgG
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.
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A00481-1 is reactive to EIF3E in Human, Mouse, Rat
Observed Molecular Weight
52 kDa
Calculated molecular weight
52.2 kDa
Background of EIF3E
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.
Antibody Validation
Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.
Application & Images
Applications
A00481-1 is guaranteed for IP, IHC, WB Boster Guarantee
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.
Western blot, 0.1-0.5μg/ml, Human, Mouse, Rat
Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Human
Immunoprecipitation, 0.5-2 μg/ml, Human
Positive Control
WB: human K562 whole cell, human 293T whole cell, human MCF-7 whole cell, human HepG2 whole cell, rat liver tissue, rat brain tissue, mouse liver tissue, mouse brain tissue
IHC: human cerebral cortex tissue, human stomach tissue
IP: HepG2 cell
Validation Images & Assay Conditions
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Western blot analysis of EIF3e using anti-EIF3e antibody (A00481-1).
Electrophoresis was performed on a 10% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human K562 whole cell lysates,
Lane 2: human 293T whole cell lysates,
Lane 3: human MCF-7 whole cell lysates,
Lane 4: human HepG2 whole cell lysates,
Lane 5: rat liver tissue lysates,
Lane 6: rat brain tissue lysates,
Lane 7: mouse liver tissue lysates,
Lane 8: mouse brain tissue 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 rabbit anti-EIF3e antigen affinity purified polyclonal antibody (A00481-1) 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-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an ECL Plus Western Blotting Substrate (Catalog # AR1196-200) 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.
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IHC analysis of EIF3e using anti-EIF3e antibody (A00481-1).
EIF3e was detected in a paraffin-embedded section of human cerebral cortex tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-EIF3e Antibody (A00481-1) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
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IHC analysis of EIF3e using anti-EIF3e antibody (A00481-1).
EIF3e was detected in a paraffin-embedded section of human stomach tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-EIF3e Antibody (A00481-1) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
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Immunoprecipitating (IP) EIF3e in HepG2 whole cell lysate.
Western blot analysis of EIF3e using anti-EIF3e antibody (A00481-1);
Lane 1: HepG2 whole cell lysates (30ug);
Lane 2: Rabbit control IgG instead of anti-EIF3e antibody in HepG2 whole cell lysate;
Lane 3: anti-EIF3e antibody (2μg) + HepG2 whole cell lysate (500μg).
After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-EIF3e antigen affinity purified polyclonal antibody (A00481-1) at a dilution of 0.5 μg/mL and probed with a goat anti-rabbit IgG-HRP secondary antibody (Light Chain). The signal is developed using ECL Plus Western Blotting Substrate (Catalog # AR1196-200). A specific band was detected for EIF3e at approximately 52 kDa. The expected band size for EIF3e is at 52 kDa.
Specific Publications For Anti-EIF3e Antibody Picoband® (A00481-1)
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1 Customer Q&As for Anti-EIF3e Antibody Picoband®
Question
We are currently using anti-EIF3e antibody A00481-1 for rat tissue, and we are well pleased with the WB results. The species of reactivity given in the datasheet says human, mouse, rat. Is it true that the antibody can work on goat tissues as well?
Verified Customer
Verified customer
Asked: 2020-03-31
Answer
The anti-EIF3e antibody (A00481-1) has not been validated for cross reactivity specifically with goat tissues, but there is a good chance of cross reactivity. We have an innovator award program that if you test this antibody and show it works in goat you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2020-03-31
