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Product Info Summary
| SKU: | A03888-2 |
|---|---|
| Size: | 100 µg/vial |
| Reactive Species: | Human, Monkey, Mouse, Rat |
| Host: | Rabbit |
| Application: | ELISA, Flow Cytometry, IF, ICC, WB |
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Product info
Product Name
Anti-P97/DAP5/EIF4G2 Antibody Picoband®
SKU/Catalog Number
A03888-2
Size
100 µg/vial
Form
Lyophilized
Description
Boster Bio Anti-P97/DAP5/EIF4G2 Antibody Picoband® catalog # A03888-2. Tested in ELISA, Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human, Mouse, Rat, Monkey. 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-P97/DAP5/EIF4G2 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A03888-2)
Host
Rabbit
Contents
Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human P97/DAP5/EIF4G2 recombinant protein (Position: R23-Q889).
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A03888-2 is reactive to EIF4G2 in Human, Monkey, Mouse, Rat
Observed Molecular Weight
97 kDa
Calculated molecular weight
102.4 kDa
Background of EIF4G2
Eukaryotic translation initiation factor 4 gamma 2 (also called p97, NAT1, and DAP-5) is a protein that in humans is encoded by the EIF4G2 gene. Translation initiation is mediated by specific recognition of the cap structure by eukaryotic translation initiation factor 4F (eIF4F), which is a cap binding protein complex that consists of three subunits: eIF4A, eIF4E and eIF4G. The protein encoded by this gene shares similarity with the C-terminal region of eIF4G that contains the binding sites for eIF4A and eIF3; eIF4G, in addition, contains a binding site for eIF4E at the N-terminus. Unlike eIF4G, which supports cap-dependent and independent translation, this gene product functions as a general repressor of translation by forming translationally inactive complexes. In vitro and in vivo studies indicate that translation of this mRNA initiates exclusively at a non-AUG (GUG) codon. Alternatively spliced transcript variants encoding different isoforms of this gene have been described.
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
A03888-2 is guaranteed for ELISA, Flow Cytometry, IF, ICC, 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, Monkey
Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human
Flow Cytometry (Fixed), 1-3 μg/1x106 cells, Human
ELISA, 0.1-0.5 μg/ml, -
Positive Control
WB: human MCF-7 whole cell, human Hela whole cell, monkey COS-7 whole cell, rat heart tissue, mouse heart tissue
ICC/IF: U87 cell
FCM: THP-1 cell
Validation Images & Assay Conditions
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Western blot analysis of P97/DAP5/EIF4G2 using anti-P97/DAP5/EIF4G2 antibody (A03888-2).
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 MCF-7 whole cell lysates,
Lane 2: human Hela whole cell lysates,
Lane 3: monkey COS-7 whole cell lysates,
Lane 4: rat heart tissue lysates,
Lane 5: mouse heart 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-P97/DAP5/EIF4G2 antigen affinity purified polyclonal antibody (Catalog # A03888-2) 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 Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for P97/DAP5/EIF4G2 at approximately 97 kDa. The expected band size for P97/DAP5/EIF4G2 is at 102 kDa.
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IF analysis of P97/DAP5/EIF4G2 using anti-P97/DAP5/EIF4G2 antibody (A03888-2).
P97/DAP5/EIF4G2 was detected in an immunocytochemical section of U87 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 μg/mL rabbit anti-P97/DAP5/EIF4G2 Antibody (A03888-2) overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG (BA1032) was used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
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Flow Cytometry analysis of THP-1 cells using anti-P97/DAP5/EIF4G2 antibody (A03888-2).
Overlay histogram showing THP-1 cells stained with A03888-2 (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 rabbit anti-P97/DAP5/EIF4G2 Antibody (A03888-2, 1 μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit 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-P97/DAP5/EIF4G2 Antibody Picoband® (A03888-2)
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