Anti-EIF5A Antibody Picoband™

Product Name

Anti-EIF5A Antibody Picoband™

View all EIF5A Antibodies

SKU/Catalog Number

A01727-3

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-EIF5A Antibody Picoband™ catalog # A01727-3. Tested in ELISA, Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human, Mouse, Rat.

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-EIF5A Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A01727-3)

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 EIF5A recombinant protein (Position: E107-K154).

*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

A01727-3 is reactive to EIF5A in Human, Mouse, Rat

Reconstitution

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

Observed Molecular Weight

18 kDa

Calculated molecular weight

57954 MW

Background of EIF5A

Eukaryotic translation initiation factor 5A-1 is a protein that in humans is encoded by the EIF5A gene. Eukaryotic initiation factor 5A (eIF5A) is an mRNA-binding protein that is involved in translation elongation and plays an important role in promoting translation of polyproline motifs. The eIF5A (eIF5A1) and eIF5A2 genes encode the two vertebrate eIF5A isoforms. While eIF5A1 is expressed constitutively in all tissues, eIF5A2 is mainly expressed in gonads. eIF5A and eIF5A2 are the only identified proteins that contain the distinctive amino acid hypusine, which is generated posttranslationally from lysine through a highly conserved polyamine metabolism pathway. eIF5A function and hypusine modification are both essential for cell proliferation, as knock down of eIF5A expression or blocking eIF5A hypusine modification suppresses cancer cell proliferation. Interestingly, eIF5A is an identified component of a tumor suppressor network of the polyamine-hypusine axis. Co-suppression of both eIF5A and adenosylmethionine decarboxylase 1 (AMD1) promotes lymphomagenesis in mice, while heterozygous deletions of the corresponding AMD1 and eIF5A genes often occur together in human lymphomas.

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.

View more details

Applications

A01727-3 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, 0.1-0.25 μg/ml, Human, Mouse, Rat
Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human
Flow Cytometry (Fixed), 1-3 μg/1x10⁶ cells, Human, Mouse
Direct ELISA, 0.1-0.5 μg/ml, Human

Positive Control

Validation Images & Assay Conditions

Click image to see more details

Figure 1. Western blot analysis of EIF5A using anti-EIF5A antibody (A01727-3).
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 Hela whole cell lysates,
Lane 2: human Jurkat whole cell lysates,
Lane 3: rat kidney tissue lysates,
Lane 4: mouse kidney 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-EIF5A antigen affinity purified polyclonal antibody (Catalog # A01727-3) at 0.25 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 EIF5A at approximately 18 kDa. The expected band size for EIF5A is at 18 kDa.

Click image to see more details

Figure 2. IF analysis of EIF5A using anti-EIF5A antibody (A01727-3).
EIF5A was detected in an immunocytochemical section of A549 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-EIF5A Antibody (A01727-3) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 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.

Click image to see more details

Figure 3. Flow Cytometry analysis of Neuro-2a cells using anti-EIF5A antibody (A01727-3).
Overlay histogram showing Neuro-2a cells stained with A01727-3 (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-EIF5A Antibody (A01727-3, 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.

Click image to see more details

Figure 4. Flow Cytometry analysis of PC-3 cells using anti-EIF5A antibody (A01727-3).
Overlay histogram showing PC-3 cells stained with A01727-3 (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-EIF5A Antibody (A01727-3, 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.

Gene/Protein Information For EIF5A (Source: Uniprot.org, NCBI)

Alternative Names

eIF4D; eIF-4D; eIF5A; EIF-5A; EIF5A1; eIF-5A1; eIF-5A-1; eIF5AI; Eukaryotic initiation factor 5A isoform 1; eukaryotic initiation factor 5A; eukaryotic translation initiation factor 5A; eukaryotic translation initiation factor 5A-1; MGC104255; MGC99547; Rev-binding Factor EIF5A EIF-5A1, eIF-4D, eIF5AI, EIF5A eukaryotic translation initiation factor 5A eukaryotic translation initiation factor 5A-1|eukaryotic initiation factor 5A|rev-binding factor

For more info on EIF5A, check out the EIF5A Infographic

We have 30,000+ of these available, one for each gene! Check them out.

In this infographic, you will see the following information for EIF5A: 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].

Take me to the EIF5A Infographic

Here are featured tools and databases that you might find useful.

• Boster's Pathways Library
• Protein Databases
• Bioscience Research Protocol Resources
• Data Processing & Analysis Software
• Photo Editing Software
• Scientific Literature Resources
• Research Paper Management Tools
• Molecular Biology Software
• Primer Design Tools
• Bioinformatics Tools
• Phylogenetic Tree Analysis