Transportin 1, also known as Karyopherin- β 2 or Importin- β 2, is part of the β-karyopherins family, which consists of importins and exportins responsible for the active transport of proteins between the nucleus and cytoplasm. Transportin 1 is composed of twenty HEAT (or a tandem repeat protein structural motif comprised of two alpha helices that end with a short loop) stacks that form a helix. In the presence of Ran-GTP, Transportin 1 undergoes a conformational change to release the cargo it’s transporting. Transportin 1 is known to bind and transport hnRNP A1, or heterogeneous ribonucleoprotein A1. hnRNPD, the ribonucleoprotein highlighted in the following articles, is a protein that binds RNA molecules with AU-rich elements (AREs). It can also function as a transcription factor when bound to DNA sequences, and has roles in mRNA biogenesis and metabolism. The following research articles use a Transportin 1 antibody in order to further understand the mechanisms of hnRNPD.
Vieira et al used a Transportin 1 antibody in their research to show that defects in hnRNPD-like protein cause limb-girdle muscular dystrophy 1G (LGM1G). In vivo analysis of hnRNPH-like protein revealed that it is associated with improper muscle development in zebrafish, which lays the groundwork for elucidating the roles of RNA binding and processing proteins in muscle development. A Transportin 1 antibody was used in immunohistochemistry (IHC) and western blot (WB) on a loss of hnRNPH function zebrafish tissue sample. After concluding that the IHC and WB staining patterns of Dystrophin, Calpain 3, Dysferlin, and Telethonin were normal in the loss of function tissue, the Transportin 1 antibody was used to look for nuclear colocalization of hnRPDL. Interestingly, IHC analysis showed that hnRPDL did in fact show nuclear colocalization that was greater than that of control tissue samples. The preliminary evidence indicates that hnRPDL alters mRNA activity, which lead to the observed muscular dystrophy effects in the study samples.
Flow (Intracellular): Transportin 1 Antibody (D45) [NB600-1397] - HeLa cells were stained with Transportin 1 NB600-1397 (blue) and a matched isotype control NBP2-27287 (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 1 ug/mL for 30 minutes at room temperature, followed by Dylight488-conjugated anti-mouse secondary antibody.
Next, Suzuki et al used a Transportin 1 antibody to further elucidate the nuclear import shuttling pathway of hnRNPD. Using a GFP-hnRNPD fusion approach in HeLa cells, the nuclear import and export of this protein was examined. To begin, a Transportin 1 antibody was used in order to determine whether the nuclear import of hnRNPD was regulated by Transportin 1. The Transportin 1 antibody was then
used in western blot to look for an interaction between Transportin 1 and nuclear localization sequence (NLS) mutants using a GST pull-down assay. As expected, NLS mutant proteins bound Transportin-1 with a strong affinity, DNS bound with a lesser affinity, and the shortest mutants acted similar to a control with zero binding affinity. These results helped Suzuki et al to identify which regions of the nucleocytoplasmic shuttling sequence were important for transport.
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