EIF5A antibody (AA 58-154)

Catalog No. ABIN968706

Product Details anti-EIF5A Antibody

Target: EIF5A (Eukaryotic Translation Initiation Factor 5A (EIF5A))

Binding Specificity: AA 58-154

Reactivity: Human, Mouse, Rat, Dog, Chicken

Host: Mouse

Clonality: Monoclonal

Conjugate: This EIF5A antibody is un-conjugated

Application: Western Blotting (WB), BioImaging (BI)

Cross-Reactivity: Chicken, Dog (Canine), Mouse (Murine), Rat (Rattus)

Characteristics: 1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
2. Please refer to us for technical protocols.
3. This antibody has been developed and certified for the bioimaging application. However, a routine bioimaging test is not performed on every lot. Researchers are encouraged to titrate the reagent for optimal performance.
4. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
5. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
6. Triton is a trademark of the Dow Chemical Company.

Purification: The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Immunogen: Human eIF-5a aa. 58-154

Clone: 26-eIF

Isotype: IgG1

Application Details

Application Notes: Bioimaging
1. Seed the cells in appropriate culture medium at ~10,000 cells per well in an 96-well Imaging Plate and culture overnight.
2. Remove the culture medium from the wells, and fix the cells by adding 100 myl of Fixation Buffer to each well. Incubate for 10 minutes at room temperature (RT).
3. Remove the fixative from the wells, and permeabilize the cells using either 90% methanol, or Triton™ X-100: a. Add 100 myl of -20°C 90% methanol to each well and incubate for 5 minutes at RT. OR b. Add 100 myl of 0.1% Triton™ X-100 to each well and incubate for 5 minutes at RT.
4. Remove the permeabilization buffer, and wash the wells twice with 100 myl of 1× PBS.
5. Remove the PBS, and block the cells by adding 100 myl of to each well. Incubate for 30 minutes at RT.
6. Remove the blocking buffer and add 50 myl of the optimally titrated primary antibody (diluted in Stain Buffer) to each well, and incubate for 1 hour at RT.
7. Remove the primary antibody, and wash the wells three times with 100 myl of 1× PBS.
8. Remove the PBS, and add the second step reagent at its optimally titrated concentration in 50 myl to each well, and incubate in the dark for 1 hour at RT.
9. Remove the second step reagent, and wash the wells three times with 100 myl of 1× PBS.
10. Remove the PBS, and counter-stain the nuclei by adding 200 myl per well of 2 myg/ml Hoechst 33342 in 1× PBS to each well at least 15 minutes before imaging.
11. View and analyze the cells on an appropriate imaging instrument.

Comment:

Related Products: ABIN968537, ABIN967389

Restrictions: For Research Use only

Handling

Format: Liquid

Concentration: 250 μg/mL

Buffer: Aqueous buffered solution containing BSA, glycerol, and ≤0.09 % sodium azide.

Preservative: Sodium azide

Precaution of Use: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Storage: -20 °C

Storage Comment: Store undiluted at -20°C.

References for anti-EIF5A antibody (ABIN968706)

Xu, Chen: "Hypusine is required for a sequence-specific interaction of eukaryotic initiation factor 5A with postsystematic evolution of ligands by exponential enrichment RNA." in: The Journal of biological chemistry, Vol. 276, Issue 4, pp. 2555-61, (2001) (PubMed).

Koettnitz, Wöhl, Kappel, Lottspeich, Hauber, Bevec: "Identification of a new member of the human eIF-5A gene family." in: Gene, Vol. 159, Issue 2, pp. 283-4, (1995) (PubMed).

Target Details for EIF5A

Target: EIF5A (Eukaryotic Translation Initiation Factor 5A (EIF5A))

Alternative Name: eIF-5a (EIF5A Products)

Synonyms: EIF-5A antibody, EIF5A1 antibody, eIF5AI antibody, EIF5A antibody, zgc:55504 antibody, zgc:77429 antibody, wu:fb05b06 antibody, wu:fc23f04 antibody, wu:fc61c10 antibody, AA410058 antibody, D19Wsu54e antibody, Eif4d antibody, Eif5a1 antibody, eIF-4D antibody, eIF-5A antibody, eIF-5A-1 antibody, eIF-5A1 antibody, zgc:77099 antibody, wu:fb17e01 antibody, wu:fb63h04 antibody, wu:fk81e04 antibody, DDBDRAFT_0191442 antibody, DDBDRAFT_0218663 antibody, DDB_0191442 antibody, DDB_0218663 antibody, 1296 antibody, CG3186 antibody, Dmel\\CG3186 antibody, anon-EST:fe2H10 antibody, eIF5A antibody, eIF5a antibody, l(2)01296 antibody, EIF-5A1 antibody, NeIF-5A3 antibody, eif5a antibody, iff-2 antibody, eukaryotic translation initiation factor 5A antibody, eukaryotic translation initiation factor 5A2 antibody, eukaryotic translation initiation factor 5a antibody, putative eukaryotic initiation factor 5a antibody, CG3186 gene product from transcript CG3186-RB antibody, eukaryotic translation initiation factor 5A-1 antibody, eukaryotic translation initiation factor 5A L homeolog antibody, EIF5A antibody, eif5a2 antibody, Eif5a antibody, eif5a antibody, tif5A antibody, Tb11.03.0410 antibody, LOC5579111 antibody, EIF5A1 antibody, eIF-5A antibody, LOC107820748 antibody, eif5a.L antibody

Background: Initiation of eukaryotic translation involves a series of reactions mediated by multiple eukaryotic initiation factors (eIFs). However, one member of this family, eIF-5a, may have multiple protein functions. eIF-5a was originally identified as an initiation factor due to its binding of ribosomes, and its stimulation of methionyl-puromycin. Regulation of initiation may not be the major function of eIF-5a, since it functions as a targeting protein for the HIV protein, Rev, and the T-cell leukemia virus type 1 protein, Rex. In Xenopus oocytes, eIF-5a interacts with the nucleoporins Nup214, Nup153, Nup98, and Nup62, and is required for Rev-NES interaction with exportin1. These interactions may be involved with eIF-5a nuclear export of Rev protein, and eIF-5a requirement for Rev-mediated viral RNA export. eIF-5a is ubiquitously expressed, and is the only known eukaryotic protein that contains the post-translationally formed hypusine residue. Modification of eIF-5a with hypusine is coupled to cell proliferation, and disruption of this residue in yeast eIF-5a is lethal. Thus, eIF-5a may be an important nuclear transport protein with conserved function in many species.

Molecular Weight: 18 kDa

Pathways: Regulation of Muscle Cell Differentiation