human 1097  (Miltenyi Biotec)

Journal: The FASEB Journal

Article Title: Prodomain processing controls BMP ‐10 bioactivity and targeting to fibrillin‐1 in latent conformation

doi: 10.1096/fj.202401694R

Figure Lengend Snippet: The processed BMP‐10 CPLX binds robustly to BMP receptors whereas the unprocessed dimer shows no receptor interaction. Sensorgrams of SPR interaction studies of soluble BMP‐10 GF, processed BMP‐10 CPLX, and unprocessed BMP‐10 dimer flowed over immobilized BMP receptors. Soluble analytes were injected onto immobilized BMPRII, ALK‐1, and ENG at concentrations ranging from 0 to 80 nM. K D s were calculated from three independent experiments ( N = 3).

Article Snippet: To assess the binding affinity of BMP‐10 processing variants to BMP receptors, the human IgG1‐Fc‐fusion ectodomains of BMPRII (#811‐BR‐100/CF, R&D Systems), ALK‐1 (#370‐AL‐100/CF, R&D Systems) as well as ENG (#1097‐EN‐025/CF, R&D Systems) were immobilized at 500 or 800 RUs on a CM5 chip via amine coupling.

Techniques: Injection

Journal: The FASEB Journal

Article Title: Prodomain processing controls BMP ‐10 bioactivity and targeting to fibrillin‐1 in latent conformation

doi: 10.1096/fj.202401694R

Figure Lengend Snippet: SPR affinity data for interactions between the immobilized BMP receptors ALK1, BMPRII, and co‐receptor ENG.

Article Snippet: To assess the binding affinity of BMP‐10 processing variants to BMP receptors, the human IgG1‐Fc‐fusion ectodomains of BMPRII (#811‐BR‐100/CF, R&D Systems), ALK‐1 (#370‐AL‐100/CF, R&D Systems) as well as ENG (#1097‐EN‐025/CF, R&D Systems) were immobilized at 500 or 800 RUs on a CM5 chip via amine coupling.

Techniques:

Journal: International Journal of Biological Sciences

Article Title: N-acetylcysteine and raloxifene boost photodynamic therapy against cutaneous squamous cell carcinoma by decreasing TGFβ1 secreted by cancer-associated fibroblasts

doi: 10.7150/ijbs.106642

Figure Lengend Snippet: Effect of CAF endoglin expression levels on TGFβ1 secretion and on the induction of resistance to PDT in A431 cells. (A) Secretion of TGFβ1 produced by T205A after treatment with increasing concentrations of recombinant endoglin. ELISA assay was performed to quantify the levels of secreted TGFβ1 to the culture medium collected 24 h after treatments. (B) Expression of endoglin analyzed by Western blot in T205A CAFs after transfection with the negative control (NC) and a siRNA targeting endoglin. Western blot images are representative of n = 3 independent experiments. Error bars denote ± S.E.M. (n = 4, one-way ANOVA: *p<0.05, **p<0.01 and ***p<0.001). (C) Secretion of TGFβ1 produced by T205A CAFs after transfection with NC or ENGsiRNA. ELISA assay was performed to quantify the levels of secreted TGFβ1 to the culture medium collected 24 h after treatment with NAC (D) Effect of the pre-treatment with NAC of transfected T205A in the CAF-derived CM on the response of bidimensional A431 cell cultures to PDT. Upper row: phase contrast images illustrating the morphological changes in A431 cells after treatments. Scale bar: 50 µm. Lower row: cell viability rates of A431 cells treated with MAL-PDT (0.5 mM of MAL and red light dose of 9.1 J·cm -2 ) in the presence of T205A CM after transfection. Error bars denote ± S.E.M. (n = 3, one-way ANOVA, statistical comparisons to Fb condition: *<0.05, **p<0.01 and ***p<0.001; multiple comparisons: #p<0.05, ##p<0.01 and ###p<0.001).

Article Snippet: Lyophilized recombinant human endoglin (R&D Systems, Minneapolis, MN, USA) was reconstituted in PBS containing 0.1% BSA to prepare a 250 μg/ml stock solution.

Techniques: Expressing, Produced, Recombinant, Enzyme-linked Immunosorbent Assay, Western Blot, Transfection, Negative Control, Derivative Assay

Journal: Electronic Journal of Biotechnology

Article Title: Isolation of adipose derived mesenchymal stem cells by using anti-CD105 VHH-magnetic beads

doi: 10.1016/j.ejbt.2023.06.004

Figure Lengend Snippet: Fig. 1. Anti-CD105 ELISA with pure EngVHH17. Purification of recombinant EngVHH17 by affinity chromatography in a Ni-NTA column. M: markers; L: E. coli lysate; E: EngVHH17. Markers Kaleidoscope Prestained Standards (BioRad).

Article Snippet: The immunization was followed by estimating the anti-T4 lysate and anti-CD105 IgG levels in serum by ELISA using recombinant CD105 (R&D Systems) and T4 protein lysate.

Techniques: Enzyme-linked Immunosorbent Assay, Recombinant, Chromatography

Journal: Electronic Journal of Biotechnology

Article Title: Isolation of adipose derived mesenchymal stem cells by using anti-CD105 VHH-magnetic beads

doi: 10.1016/j.ejbt.2023.06.004

Figure Lengend Snippet: Fig. 2. (a) A saturation binding curve with a concentration range of CD105 by EngVHH17-ELISA. Each point is the mean of OD450 values of duplicate assay. One-site hyperbole fit (R2 = 0.99). (b) The Scatchard plot of the binding EngVHH17 to CD105. The Scachard equation [B]/[F] = -2.56[B] + 2.76 was fitted by a linear regression (R2 = 0.98), a dissociation constant value (Kd) of 3.9 10–10 M was estimated for the binding of CD105 to EngVHH17.

Article Snippet: The immunization was followed by estimating the anti-T4 lysate and anti-CD105 IgG levels in serum by ELISA using recombinant CD105 (R&D Systems) and T4 protein lysate.

Techniques: Binding Assay, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Electronic Journal of Biotechnology

Article Title: Isolation of adipose derived mesenchymal stem cells by using anti-CD105 VHH-magnetic beads

doi: 10.1016/j.ejbt.2023.06.004

Figure Lengend Snippet: Fig. 3. Western blot with T24 cell lysate and EngVHH17. (a) T24 lysate resolved in 15% SDS-PAGE. M: molecular weight markers; L: T24 cell lysate. (b) Western blot with EngVHH17 showing a 95 kDa band corresponding to immune detection of the CD105 monomer present in the T24 cell lysate.

Article Snippet: The immunization was followed by estimating the anti-T4 lysate and anti-CD105 IgG levels in serum by ELISA using recombinant CD105 (R&D Systems) and T4 protein lysate.

Techniques: Western Blot, SDS Page, Molecular Weight

Journal: Electronic Journal of Biotechnology

Article Title: Isolation of adipose derived mesenchymal stem cells by using anti-CD105 VHH-magnetic beads

doi: 10.1016/j.ejbt.2023.06.004

Figure Lengend Snippet: Fig. 4. Visualization of CD105 on the cell surface of differentiated THP1 cells. Human THP-1 cells were treated with 65 nM phorbol 12-myristate 13-acetate (PMA) for 72 h. THP1 (2.5 104 cells/well) were fixed and CD105 immunolabeled by using anti-CD105 IgG2a mAb and secondary antibody Alexa Fluor TM 594 labeled goat anti-mouse IgG2a, or anti-CD105 EngVHH17 and FITC anti-6 His Tag mouse mAb, and DAPI. Undifferentiated THP1 cells do not show immunofluorescence labeling with anti-CD105 mouse IgG2a mAb (panel a); anti-CD105 EngVHH17 (panel b); PMA-treated THP1 cells for 72 h showed immunofluorescence labeling on the cell surface with anti-CD105 mouse IgG2a mAb (panel c); anti-CD105 EngVHH17 (panel d); Nuclear DNA of THP1 cells was fluorescence stained with DAPI (blue). Images were taken on a Zeiss LSM 880 confocal microscope at 20 magnification.

Article Snippet: The immunization was followed by estimating the anti-T4 lysate and anti-CD105 IgG levels in serum by ELISA using recombinant CD105 (R&D Systems) and T4 protein lysate.

Techniques: Immunolabeling, Labeling, Staining, Microscopy