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Product Info Summary
| SKU: | M00481-1 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Mouse |
| Application: | Flow Cytometry, WB |
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Product info
Product Name
Anti-EIF3e Antibody Picoband® (monoclonal, 10F5H6)
SKU/Catalog Number
M00481-1
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-EIF3e Antibody Picoband® (monoclonal, 10F5H6) catalog # M00481-1. Tested in Flow Cytometry, 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
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.
Cite This Product
Anti-EIF3e Antibody Picoband® (monoclonal, 10F5H6) (Boster Biological Technology, Pleasanton CA, USA, Catalog # M00481-1)
Host
Mouse
Contents
Each vial contains 4 mg Trehalose, 0.9 mg NaCl and 0.2 mg Na2HPO4.
Clonality
Monoclonal
Clone Number
10F5H6
Isotype
Mouse IgG2b
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
M00481-1 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
M00481-1 is guaranteed for Flow Cytometry, 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, 0.25-0.5 μg/ml, Human, Mouse, Rat
Flow Cytometry (Fixed), 1-3 μg/1x106 cells, Human
Positive Control
WB: human Raji whole cell, human K562 whole cell, rat thymus tissue, mouse thymus tissue, mouse ANA-1 whole cell, mouse SP2/0 whole cell
FCM: U20S cell
Validation Images & Assay Conditions
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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.
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Flow Cytometry analysis of U20S cells using anti-EIF3E antibody (M00481-1).
Overlay histogram showing U20S cells stained with M00481-1 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. 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 without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Specific Publications For Anti-EIF3e Antibody Picoband® (monoclonal, 10F5H6) (M00481-1)
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