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Production and Characterization of Monoclonal Antibodies aganist Heat Shock Protein 70
Kang, Ho Sung,Park, Young Chul,Han, Song Iy,Kim, Young Hee,Moon, Dae Yeon,Lee, Chu,Kim, Byeong Gee,Hah, Jae Chung,Kim, Chong-Rak,Kim, Han Do 부산대학교 기초과학연구소 1995 부산대학교 기초과학연구소 연구논문집 Vol.15 No.-
Heat shock protein 70(hsp70, also known as hsp72/hsp73) is known to function as a molecular chaperone that regulates in vivo folding and assembly of proteins. However, its exact action mechanism remains to be elucidated. In this study, in order to facilitate the characterization of hsp70 function, monoclonal antibodies against hsp70 and BiP/grp78 were produced by the murine hybridoma technique. For the production of MAb's, hsp70 and BiP/grp78 were purified from human placenta by DEAE-cellulose ion exchange and ATP-agarose affinity chromatography and immunized into Balb/c mice and then the spleen cells from the mice were fused with NS1 myeloma cells, The hybridoma cells were screened by enzyme-linked immunosorbent assay and subcloned by limiting dilution and successively in semi-solid agarose. Here we report the production of 16 positive monoclonal hybridoma cells which all recognized inducible hsp72 as well as constitutively expressed hsp73. Most of the monoclonal antibodies showed the cross-reactivities with hsp70 from human, rabbit, mouse, chick, plant, frog, fish and Drosophila but not with that from E. coli.
Lee, Young-Rae,Noh, Eun-Mi,Kwon, Kang-Beom,Lee, Yun-Sik,Chu, Jong Phil,Kim, Eun Jib,Park, Young-Seok,Kim, Byeong-Soo,Kim, Jong-Suk D.A. Spandidos 2009 INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE Vol.23 No.5
<P>Ultraviolet (UV)B irradiation induces the production of matrix metalloproteinases (MMPs), which are responsible for the degradation of collagenous extracellular matrix in connective tissues, causing skin photoaging. Although Radix clematidis is commonly used in Chinese medicine for the treatment of arthralgia, the anti-skin photoaging effects of Radix clematidis have not yet been reported. In the present study, we investigated the inhibitory effects of Radix clematidis extract (RCE) on MMP-1 and -3 expression of human dermal fibroblast cells via various in vitro experiments and elucidated the pathways of inhibition. Western blot analysis and real-time PCR revealed RCE inhibited UVB-induced MMP-1 and -3 expressions in a dose-dependent manner. UVB strongly activated nuclear factor-kappaB (NF-kappaB) activity, which was determined by IkappaBalpha degradation, nuclear localization of p50 and p65 subunit, and NF-kappaB binding activity. However, UVB-induced NF-kappaB activation was completely blocked by RCE pretreatment. These findings suggest that RCE prevents UVB-induced MMP expression through inhibition of NF-kappaB activation. In conclusion, RCE is a potential agent for the prevention and treatment of skin photoaging.</P>