fibrillin 1  (Bioss)

Journal: Cells

Article Title: Development and Characterization of Alkaline Phosphatase-Positive Human Umbilical Cord Perivascular Cells

doi: 10.3390/cells10113011

Figure Lengend Snippet: Morphological observation of HUCPVCs by fluorescent immunostaining using elastin and fibrillin-1 antibodies. Fluorescence detection using Alexa Fluor 546-labeled elastin (ELN) and Alexa Fluor 488-labeled fibrillin-1 (FBN-1) into V-HUCPVCs, VL-HUCPVCs, and ALP(+)-HUCPVCs. Each image was compared to HUCPVCs cultured in the absence of VD, LDN, and TGF-β1 (Control). Cells with co-existing elastin and fibrillin-1 were detected in the merged image, which displayed light-red fluorescence (Merge). Elastin (red) and fibrillin-1 (green) in fixed HUCPVCs were detected using human elastin and fibrillin-1 antibodies at a dilution of 1:200, respectively. For secondary antibody application, Alexa Fluor 546-conjugated rabbit anti-mouse IgG (H+L) antibody for elastin and an Alexa Fluor 488 goat anti-rabbit IgG (H+L) antibody for fibrillin-1 were used at 1:500. DAPI was used to stain the cell nuclei (blue) at a concentration of 2 μg/mL (scale bar, 50 μm).

Article Snippet: For the primary antibody application, an anti-elastin monoclonal antibody (#ab9519, Abcam, Cambridge, UK), anti-fibrillin-1 polyclonal antibody (#BS-1157R, Bioss Antibodies Inc., Woburn, MA, USA), anti-osteopontin polyclonal antibody (#AF808, R&D Systems, Minneapolis, MN, USA), and anti α-SMA antibody (#23081-1-AP, Proteintech Japan, Tokyo, Japan) were used at 1:200 or 1:300 dilutions, and the cells were incubated in the above antibody mixture overnight at 4 °C.

Techniques: Immunostaining, Fluorescence, Labeling, Cell Culture, Staining, Concentration Assay

Journal: Cells

Article Title: Development and Characterization of Alkaline Phosphatase-Positive Human Umbilical Cord Perivascular Cells

doi: 10.3390/cells10113011

Figure Lengend Snippet: Quantitative PCR (qPCR) analysis of tissue-specific differentiation marker genes for V-HUCPVCs, VL-HUCPVCs, and ALP(+)-HUCPVCs on day 7 after the culture. Control: HUCPVCs cultured in the absence of VD, LDN, and TGF-β1. COL I , collagen type I alpha 1 chain; OPN , osteopontin; RUNX2 , runt-related transcription factor 2; OC , osteocalcin; ELN , elastin; FBN-1 , fibrillin-1; MYOD1 , myoblast determination protein 1; MYF5 , myogenic factor 5; POSTN , periostin; PLAP1 , periodontal ligament associated protein-1; CD73 , CD90 , CD105 , cluster of differentiation 73, 90, 105; α-SMA , alpha-smooth muscle actin. ND, not determined. Each mRNA expression value was normalized to that of the reference gene GAPDH , and the relative quantification data for COL I , OPN , RUNX2 , OC , ELN , FBN-1 , MYOD1 , MYF5 , POSTN , PLAP1 , CD90 , α-SMA , CD73 , and CD105 in HUCPVC were generated on the basis of a mathematical model for relative quantification in a qPCR system ( n = 6). All values are presented as the mean ± SEM (* p < 0.05, Steel–Dwass test).

Article Snippet: For the primary antibody application, an anti-elastin monoclonal antibody (#ab9519, Abcam, Cambridge, UK), anti-fibrillin-1 polyclonal antibody (#BS-1157R, Bioss Antibodies Inc., Woburn, MA, USA), anti-osteopontin polyclonal antibody (#AF808, R&D Systems, Minneapolis, MN, USA), and anti α-SMA antibody (#23081-1-AP, Proteintech Japan, Tokyo, Japan) were used at 1:200 or 1:300 dilutions, and the cells were incubated in the above antibody mixture overnight at 4 °C.

Techniques: Real-time Polymerase Chain Reaction, Marker, Cell Culture, Expressing, Generated