Rabbit Anti-EIF5A Recombinant Antibody (
5E1) (V2LY-0725-LY1491)

Name: Rabbit Anti-EIF5A Recombinant Antibody (5E1)
SKU: V2LY-0725-LY1491
Rating: 5 (1 reviews)

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Tested Data

Rabbit Anti-EIF5A Recombinant Antibody (5E1) (V2LY-0725-LY1491) in WB

Western Blot
Positive WB detected in: A2780 whole cell lysate, Hela whole cell lysate, MCF-7 whole cell lysate, Jurkat whole cell lysate, Raji whole cell lysate
All lanes: EIF5A antibody at 1:2000
Secondary
Goat polyclonal to rabbit IgG at 1/50000 dilution
Predicted band size: 17, 21 kDa
Observed band size: 18 kDa

Rabbit Anti-EIF5A Recombinant Antibody (5E1) (V2LY-0725-LY1491) in IHC

IHC image of V2LY-0725-LY1491 diluted at 1:100 and staining in paraffin-embedded human colon cancer performed on a Leica BondTM system. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4°C overnight. The primary is detected by a Goat anti-rabbit IgG polymer labeled by HRP and visualized using 0.05% DAB.

Rabbit Anti-EIF5A Recombinant Antibody (5E1) (V2LY-0725-LY1491) in IF

Immunofluorescence staining of Hela Cells with V2LY-0725-LY1491 at 1:50, counter-stained with DAPI. The cells were fixed in 4% formaldehyde, permeated by 0.2% TritonX-100, and blocked in 10% normal Goat Serum. The cells were then incubated with the antibody overnight at 4°C. Nuclear DNA was labeled in blue with DAPI. The secondary antibody was FITC-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L).

Rabbit Anti-EIF5A Recombinant Antibody (5E1) (V2LY-0725-LY1491) in FC

Overlay histogram showing Jurkat cells stained with V2LY-0725-LY1491 (red line) at 1:50. The cells were fixed with 70% Ethylalcohol (18h) and then incubated in 10% normal goat serum to block non-specific protein-protein interactions followedby the antibody (1µg/1*106 cells) for 1 h at 4°C.The secondary antibody used was FITC-conjugated goat anti-rabbit IgG (H+L) at 1/200 dilution for 30min at 4°C. Control antibody (green line) was Rabbit IgG (1µg/1*106 cells) used under the same conditions. Acquisition of>10,000 events was performed.

Datasheet

SDS

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Datasheet Target References Q & As Review & reward Protocols Associated Products

Basic Information

Host Animal

Rabbit

Clone

5E1

Application

ELISA, WB, IHC, IF, FC

Immunogen

A synthesized peptide derived from human eIF5A.

Host Species

Rabbit

Specificity

Human

Antibody Isotype

IgG

Clonality

Monoclonal Antibody

Application Notes

Application	Note
WB	1:500-1:5,000
IHC	1:50-1:200
IF	1:20-1:200
FC	1:20-1:200

Formulations & Storage [For reference only, actual COA shall prevail!]

Format

Liquid

Buffer

PBS, glycerol

Preservative

Sodium azide

Concentration

Batch dependent

Purity

> 95% Purity determined by SDS-PAGE.

Storage

Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C long term. Avoid repeated freezethaw cycles.

More Infomation

Target

Full Name

Eukaryotic Translation Initiation Factor 5A

Entrez Gene ID

1984

UniProt ID

P63241

Research Area

mRNA-binding protein involved in translation elongation. Has an important function at the level of mRNA turnover, probably acting downstream of decapping. Involved in actin dynamics and cell cycle progression, mRNA decay and probably in a pathway involved in stress response and maintenance of cell wall integrity. With syntenin SDCBP, functions as a regulator of p53/TP53 and p53/TP53-dependent apoptosis. Regulates also TNF-alpha-mediated apoptosis. Mediates effects of polyamines on neuronal process extension and survival. May play an important role in brain development and function, and in skeletal muscle stem cell differentiation. Also described as a cellular cofactor of human T-cell leukemia virus type I (HTLV-1) Rex protein and of human immunodeficiency virus type 1 (HIV-1) Rev protein, essential for mRNA export of retroviral transcripts.

Biological Process

Cellular response to virus Source: UniProtKB
mRNA transport Source: UniProtKB-KW
Positive regulation of intrinsic apoptotic signaling pathway by p53 class mediator Source: UniProtKB
Positive regulation of transcription by RNA polymerase II Source: UniProtKB
Positive regulation of translational elongation Source: UniProtKB
Positive regulation of translational termination Source: InterPro
Protein transport Source: UniProtKB-KW
Tumor necrosis factor-mediated signaling pathway Source: UniProtKB

Cellular Location

Endoplasmic reticulum membrane; Nucleus; Nuclear pore complex; Cytoplasm. Hypusine modification promotes the nuclear export and cytoplasmic localization and there was a dynamic shift in the localization from predominantly cytoplasmic to primarily nuclear under apoptotic inducing conditions.

PTM

Acetylated. Deacetylated by SIRT2.
eIF-5A seems to be the only eukaryotic protein to have a hypusine residue which is a post-translational modification of a lysine by the addition of a butylamino group (from spermidine).

Barba-Aliaga, M., & Alepuz, P. (2022). Role of eIF5A in Mitochondrial Function. International Journal of Molecular Sciences, 23(3), 1284.

Li, H., Wu, B. K., Kanchwala, M., Cai, J., Wang, L., Xing, C., ... & Pan, D. (2022). YAP/TAZ drives cell proliferation and tumour growth via a polyamine–eIF5A hypusination–LSD1 axis. Nature cell biology, 24(3), 373-383.

Faundes, V., Jennings, M. D., Crilly, S., Legraie, S., Withers, S. E., Cuvertino, S., ... & Banka, S. (2021). Impaired eIF5A function causes a Mendelian disorder that is partially rescued in model systems by spermidine. Nature communications, 12(1), 1-13.

Park, M. H., Kar, R. K., Banka, S., Ziegler, A., & Chung, W. K. (2021). Post-translational formation of hypusine in eIF5A: Implications in human neurodevelopment. Amino acids, 1-15.

Bourourou, M., Gouix, E., Melis, N., Friard, J., Heurteaux, C., Tauc, M., & Blondeau, N. (2021). Inhibition of eIF5A hypusination pathway as a new pharmacological target for stroke therapy. Journal of Cerebral Blood Flow & Metabolism, 41(5), 1080-1090.

Cougnon, M., Carcy, R., Melis, N., Rubera, I., Duranton, C., Dumas, K., ... & Pisani, D. F. (2021). Inhibition of eIF5A hypusination reprogrammes metabolism and glucose handling in mouse kidney. Cell Death & Disease, 12(4), 1-14.

Coni, S., Serrao, S. M., Yurtsever, Z. N., Di Magno, L., Bordone, R., Bertani, C., ... & Canettieri, G. (2020). Blockade of EIF5A hypusination limits colorectal cancer growth by inhibiting MYC elongation. Cell death & disease, 11(12), 1-14.

Zhang, H., Alsaleh, G., Feltham, J., Sun, Y., Napolitano, G., Riffelmacher, T., ... & Simon, A. K. (2019). Polyamines control eIF5A hypusination, TFEB translation, and autophagy to reverse B cell senescence. Molecular cell, 76(1), 110-125.

Manjunath, H., Zhang, H., Rehfeld, F., Han, J., Chang, T. C., & Mendell, J. T. (2019). Suppression of ribosomal pausing by eIF5A is necessary to maintain the fidelity of start codon selection. Cell reports, 29(10), 3134-3146.

Turpaev, K. T. (2018). Translation factor eIF5A, modification with hypusine and role in regulation of gene expression. eIF5A as a target for pharmacological interventions. Biochemistry (Moscow), 83(8), 863-873.

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Please try the standard protocols which include: protocols, troubleshooting and guide.

Enzyme-linked Immunosorbent Assay (ELISA)
Flow Cytometry
Immunofluorescence (IF)
Immunohistochemistry (IHC)
Immunoprecipitation (IP)
Western Blot (WB)
Enzyme Linked Immunospot (ELISpot)
Proteogenomic
Other Protocols

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For research use only. Not intended for any clinical use.

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