Anti hnRNP-U / SAF-A polyclonal antibody


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Background

Heterogeneous nuclear ribonucleoprotein U (hnRNP-U, also known as scaffold attachment factor A, SAF-A) is a nuclear matrix-associated protein that interacts with chromosomal DNA. hnRNP-U specifically binds to scaffold/matrix attachment region of DNA and could thus be involved in higher order chromatin structure. hnRNP-U is also a RNA binding protein and forms complexes with heterogeneous nuclear RNA (hnRNA) and plays an important role in pre-mRNA processing and transport.
HnRNP-U is reported to interact with necdin, a growth suppressor that is expressed in terminally differentiated neurons and skeletal muscle cells. It has been shown that hnRNP-U recruits necdin to the nuclear matrix where they form a stable complex. It is suggested that necdin suppresses cell proliferation through its interaction with hnRNP-U in the specific subnuclear structure.

Data Link: Swiss-Prot Q8VEK3 (mouse), Q00839 (human)

Application

1. Western blotting (dilution: 1/3,000-1/1,000)
2. Immunocytochemistry (dilution: 1/1,000-1/500)
3. Immunoprecipitation 

BAM_70415EX_01.jpg
< Immunoblotting >
Specificity of anti-hnRNP-U antibody, HUT.
Cell lysates were prepared from SAOS-2 cells transfected with pRc/CMV vectors (pRc) or pRc/CMV vectors expressing Myc-tagged hnRNP-U (Myc-UF). Exogenous Myc-tagged hnRNP-U (Myc-U) and endogenous hnRNP-U (U) proteins were detected by immunoblotting with anti-Myc antibody (αMyc) or HUT (αU).
This antibody recognized exogenous Myc-tagged hnRNP-U and endogenous ~120 kDa hnRNP-U proteins in SAOS-2 cells.

BAM_70415EX_02.jpg
< Immunocytochemistry >
Mouse P19 neurons were labeled with anti-necdin antibody (Ndn) (a) or with HUT for hnRNP-U (U) (c) in combination with anti-neuronal marker, MAP2, antibody for MAP2 (b, d). The nuclear matrix was prepared in situ and labeled for necdin (Ndn) (e), hnRNP-U (U) (f), and a nuclear matrix marker, lamin B (g).
Both necdin and hnRNP-U were localized to the nuclei of differentiated neurons, which express the neuronal marker MAP2 (a-d). Necdin was also distributed in the neuronal cytoplasm (a).
The immunocytochemical analysis of in situ extracted nuclear matrix revealed that both necdin and hnRNP-U were concentrated in intranuclear speckles throughout the nucleoplasm (e, f). Lamin B, a nuclear matrix marker, was localized to the nuclear lamina (g). These results suggest that both necdin and hnRNP-U are associated with the nuclear matrix of neurons

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