The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/1000 - 1/10000. Detects a band of approximately 17 kDa (predicted molecular weight: 17 kDa).
1/10 - 1/100.
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
1/10 - 1/100.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
1/100 - 1/250.
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. Functions as a regulator of 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.
Expressed in ovarian and colorectal cancer cell lines (at protein level). Highly expressed in testis. Overexpressed in some cancer cells.
Belongs to the eIF-5A family.
eIF-5A seems to be the only eukaryotic protein to have an hypusine residue which is a post-translational modification of a lysine by the addition of a butylamino group (from spermidine).
Cytoplasm. Nucleus. Endoplasmic reticulum membrane. Nucleus > nuclear pore complex. 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.
Overlay histogram showing HeLa cells stained with ab126735 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab126735, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
Western blot - Anti-eIF5A2 antibody [EPR7411] (ab126735)
All lanes : Anti-eIF5A2 antibody [EPR7411] (ab126735) at 1/1000 dilution
Lane 1 : 293T cell lysate Lane 2 : HeLa cell lysate Lane 3 : A431 cell lysate Lane 4 : CaCo 2 cell lysate Lane 5 : HepG2 cell lysate
Lysates/proteins at 10 µg per lane.
Secondary All lanes : Goat-anti-rabbit HRP at 1/2000 dilution
Meng QB et al.
Overexpression of eukaryotic translation initiation factor 5A2 (EIF5A2) correlates with cell aggressiveness and poor survival in gastric cancer.
PLoS One10:e0119229 (2015).
Read more (PubMed: 25793713) »
Qin X et al. Inhibition of eIF5A results in aberrant uterine natural killer cell function and embryo loss in mice. Am J Reprod Immunol71:229-40 (2014).
Read more (PubMed: 24382123) »