Anti-P97/DAP5/EIF4G2 Antibody Picoband™

Product Name

Anti-P97/DAP5/EIF4G2 Antibody Picoband™

View all DAP5 Antibodies

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.

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).

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross-reactivity with other proteins.

Reactive Species

A03888-2 is reactive to EIF4G2 in Human, Monkey, Mouse, Rat

Applications

A03888-2 is guaranteed for ELISA, Flow Cytometry, IF, ICC, WB Boster Guarantee

Observed Molecular Weight

97 kDa

Calculated molecular weight

102.362kDa

Background of DAP5

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.

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Reconsitution

Adding 0.2 ml of distilled water will yield a concentration of 500 µg/ml.

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, 1-3 µg/1x10⁶ cells, Human
Direct ELISA, 0.1-0.5 µg/ml, Human

Validation Images & Assay Conditions

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Figure 1. 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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Figure 2. 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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Figure 3. 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). 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 (Red line) was also used as a control.

Gene/Protein Information For EIF4G2 (Source: Uniprot.org, NCBI)

Alternative Names

aging-associated protein 1; DAP-5; DAP5AAG1; Death-associated protein 5; eIF4G 2; eIF-4G 2; eIF-4-gamma 2; eukaryotic translation initiation factor 4 gamma 2; eukaryotic translation initiation factor 4 gamma, 2; eukaryotic translation initiation factor 4G-like 1; FLJ41344; NAT1; p97 EIF4G2 AAG1, DAP5, NAT1, P97 eukaryotic translation initiation factor 4 gamma 2 eukaryotic translation initiation factor 4 gamma 2|DAP-5|aging-associated protein 1|death-associated protein 5|eIF-4-gamma 2|eIF-4G 2|eIF4G 2|eukaryotic translation initiation factor 4G-like 1

For more info on EIF4G2, check out the EIF4G2 Infographic

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In this infographic, you will see the following information for EIF4G2: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].

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