Anti elastin antibody

The plant extracts from pomegranate (Punica granatum L.), osmanthus (Osmanthus fragrans Lour.) and olive (Olea europaea L.) were prepared using the dry plants powders dissolved in an aqueous solution under stirring and filtered. The active molecules, punicalagin, verbascoside and hydroxytyrosol corresponding to pomegranate, osmanthus and olive, respectively, were detected using high performance liquid chromatography (HPLC) analysis according to the Pharmacopoeia of the People’s Republic of China (version 2010). The standard substances of punicalagin, verbascoside and hydroxytyrosol were dissolved in dimethyl sulfoxide (DMSO) and then diluted into the cell culture medium.
Rabbit polyclonal anti-Collagen I (Novotec, France), Rabbit polyclonal anti-Elastin antibody (Abcam, UK), Mouse polyclonal anti-Melan A (Novotec, France), Rabbit polyclonal anti-PMEL (Sigma, USA), Rabbit polyclonal anti-Malondialdehyde (Abcam, UK) and Alexa-Fluor 568 anti-rabbit secondary antibody (Thermo Fisher Scientific, USA) were used for immunofluorescence staining.

In addition to the anti-LTBP2 antibody and anti-αSMA antibody, for human lung sections, anti-CD45 antibody (1:100; Dako, U.S.A.), anti-CD324 antibody (1:20; Dako, U.S.A.), anti-CD31 antibody (1:100; Dako, U.S.A.), anti-podoplanin antibody (1:200; Dako, U.S.A.), and anti-h-caldesmon antibody (1:5; MyBioSource, Canada), anti-elastin antibody (1:500; Abcam, U.S.A.), and anti-fibrillin-1 antibody (1:200; EMD Millipore, U.S.A.) were also used as primary antibodies. The sections were incubated with Hoechst 33342 (1:1000; Sigma, U.S.A.) with Alexa fluor conjugated secondary antibodies for 30 min. The sections were visualized using a confocal microscope (Leica, U.S.A.). In the present study, human lung myofibroblasts in immunofluorescence images were defined as single cells with positivity for αSMA and negativity for lineage markers: CD45 (blood cells), CD324 (epithelial cells), CD31 (vascular endothelial cells), podoplanin (lymphatic endothelial cells), and h-caldesmon (smooth muscle cells).