Human/Mouse/Rat eIF5A Antibody

Catalog #: AF7558 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
AF7558
AF7558-SP

Detection of Human, Mouse, and Rat eIF5A by Western Blot.

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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human/Mouse/Rat eIF5A Antibody Summary

Species Reactivity

Human, Mouse, Rat

Specificity

Detects human, mouse and rat eIF5A in Western blots.

Source

Polyclonal Sheep IgG

Purification

Antigen Affinity-purified

Immunogen

E. coli-derived recombinant human eIF5A
Lys85-Lys154
Accession # P63241

Formulation

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.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Western Blot

1 µg/mL

See below

Immunohistochemistry

5-25 µg/mL

See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot

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Detection of Human, Mouse, and Rat eIF5A by Western Blot. 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.

Immunohistochemistry

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eIF5A in Mouse Embryo. 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.

Immunohistochemistry

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eIF5A in Human Liver. 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.

Immunohistochemistry

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eIF5A in Rat Brain. 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.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Sterile PBS to a final concentration of 0.2 mg/mL.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: eIF5A

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.

Long Name

Eukaryotic Translation Initiation Factor 5A

Entrez Gene IDs

1984 (Human); 276770 (Mouse); 287444 (Rat)

Alternate 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