eIF5A Antibody [Unconjugated]

Western blot shows lysates of 786-O human renal cell adenocarcinoma cell line, C2C12 mouse myoblast cell line, and NR8383 rat alveolar macrophage cell line. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human/Mouse/Rat eIF5A Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7558) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for eIF5A at approximately 18 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

eIF5A was detected in immersion fixed frozen sections of mouse embryo (13 d.p.c.) using Sheep Anti-Human/Mouse/Rat eIF5A Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7558) at 5 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Sheep IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC006). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to developing central nervous system. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

eIF5A was detected in immersion fixed paraffin-embedded sections of human liver using Sheep Anti-Human/Mouse/Rat eIF5A Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7558) at 1.7 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Sheep IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC006). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in hepatocytes. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

eIF5A was detected in perfusion fixed frozen sections of rat brain (brainstem medulla) using Sheep Anti-Human/Mouse/Rat eIF5A Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7558) at 5 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Sheep IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC006). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in neurons. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

• Reactivity: Hu, Mu, Rt
• Applications: WB, IHC
• Clonality: Polyclonal
• Host: Sheep
• Conjugate: Unconjugated
• Concentration: LYOPH

eIF5A Antibody [Unconjugated] Summary

• Immunogen: E. coli-derived recombinant human eIF5A
  Lys85-Lys154
  Accession # P63241
• Specificity: Detects human, mouse and rat eIF5A in Western blots.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Sheep
• Gene: EIF5A
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Immunohistochemistry 5-25 ug/mL
  • Western Blot 1 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  12 months from date of receipt, -20 to -70 degreesC as supplied. 1 month, 2 to 8 degreesC under sterile conditions after reconstitution. 6 months, -20 to -70 degreesC under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Sterile PBS to a final concentration of 0.2 mg/mL.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for eIF5A Antibody [Unconjugated]

• 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

Background

eIF5A (Elongation initiation factor 5A; also eIF5A1, Rev-binding factor and eIF-4D) is a 16-19 kDa member of the eIF5A family of molecules. It is ubiquitously expressed, being found in all human cells examined. eIF5A acts as a stabilizer of RNA transcripts and acts as a positive modulator of p53-mediated apoptosis. It also may be involved in the formation of the first peptide bond during select protein synthesis, and is known to be essential for cell cycle progression through G1/S. The human eIF5A precursor is 154 amino acids (aa) in length. It contains a 71 aa DOHH (deoxyhypusine hydroxylase)-binding site (aa 20-90) and a 69 aa RNA-binding domain (aa 84-152). eIF5A undergoes a posttranslational modification that converts the precursor peptide into a mature molecule. This modification occurs on Lys50, and involves the transfer and hydroxylation of an aminobutyl sequence from spermidine, creating a unique amino acid termed hypusine (hydroxyputrisine lysine). When hypusine is unacetylated, eIF5A is active; when acetylated, eIF5A is inactive. Nonhypusinated eIF5A is found in both cytoplasm and nucleus, while hypusinated eIF5A is targeted to the cytoplasm. There is one isoform variant that shows a 32 aa substitution for aa 1-32. Over aa 85-154, human and mouse are identical in aa sequence. Over the same aa sequence, human eIF5A (on Chr 17p13) also shares 98% and 69% aa identity with human eIF5A1-Like (on Chr 10q22) and human eiF5A2 (on Chr 3q26), respectively.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Bioinformatics

• Gene Symbol: EIF5A
• Uniprot:
  • P63241(Human)