primary antibody for osteocalcin  (Proteintech)

Journal: Bioactive Materials

Article Title: Ribbon-shaped microgels as bioinks for 3D bioprinting of anisotropic tissue structures

doi: 10.1016/j.bioactmat.2025.12.040

Figure Lengend Snippet: μRB bioinks modulate MSC morphology, osteogenesis and bone formation in a stiffness-dependent manner. (A) Live/dead staining of MSCs after extrusion (Day 0) or after 28 days of culture in osteogenic medium. Green: live cells, Red: dead cells. (B) Metabolic activity of MSCs measured by PrestoBlue assay at day 0 and day 14 after bioprinting (n = 6 per group). (C) DNA content per scaffold measured by PicoGreen assay at day 28 (n = 3 per group). (D) Alizarin red S (ARS) staining for mineralized bone matrix, (E) Aniline Blue staining for total collagen, and (F) immunostaining of Osteocalcin (OCN), a mature bone marker, at day 14 and day 21 of osteogenesis (n = 4 per group). (G–I) Quantification of ARS and Aniline Blue percent positive area and OCN mean fluorescence intensity (MFI). Scale bar = 100 μm. Values are presented as mean ± S.D. and p-values were determined by one-way analysis of variance (ANOVA) with Tukey's multiple comparisons test; ∗p ≤ 0.05, ∗∗p ≤ 0.01, ∗∗∗p ≤ 0.005, ∗∗∗∗p ≤ 0.001.

Article Snippet: Primary antibody for osteocalcin (1:200, 23418-1-AP, Proteintech), GFP (1:100, 50430-2-AP, Proteintech) was diluted in 1 % BSA with 0.1 % Triton X-100 and incubated overnight at 4 °C.

Techniques: Staining, Activity Assay, Prestoblue Assay, Picogreen Assay, Immunostaining, Marker, Fluorescence

Journal: Bioactive Materials

Article Title: A large puncture closer of aortic wall by multi-memory actions with thrombo-hemodynamic control

doi: 10.1016/j.bioactmat.2025.12.042

Figure Lengend Snippet: 18 Fr puncture of hemostasis in porcine aorta using VWP by validating the memory programming effect of each part. a, As a challenging model for application of large-diameter catheters, i) an 18 Fr (6 mm) puncture is created into the porcine thoracic aorta (diameter: 10 mm) so that the size-matched VWP is deployed, followed by measuring proximal and distal blood pressure. ii) The experimental groups are designed first to exam the memory programming effect of collaboration between Ring squeezing with Body expansion on self-locking (SL) to enable efficient hemostasis. Next, the effect of Wing shape recovery from curve to flat is examined on hemodynamic control (HC) in cooperation with the actions of Body and Ring to handle hemostasis. No recovery of Wing shape is expected to induce excessive thrombosis. iii) Four experimental groups are established using a total of 12 pigs (N = 12) with immediate sacrifice following deployment (N = 3 each). Group 1 [SL(−) HC(−)] represents no memory programming. Group 2 [SL(+) w/flat Wing] has the effects of Body and Ring actions except the hemostatic sealing by keeping the flat Wing. Group 3 [SL(+) HC(+)] possesses the complete memory effects of the three parts. Group 4 [SL(+) w/bump Wing] is expected to have excessive thrombosis because of no shape recovery from the curved Wing while maintaining the memory actions of Body and Wing. b, Each group is visually explained in the illustrations. c , In VWP actions, (left) the bleeding condition preserves the normal sinusoidal waveform of high proximal pressure (green) in contrast to the disturbed waveform of low distal pressure (red). (middle) Hemostatic closure results in similar high sinusoidal waveform at both pressure sites. (right) Excessive thrombosis does not disturb the waveform, but the distal pressure level becomes lower than the proximal one. d, When reperfusion starts by removing the clamp post-deployment (blue), only Group 3 [SL(+) HC(+)] reaches the hemostatic closure, as evidenced by flow stabilization (red) with a 5 s plateau at both pressure sites. Group 4 [SL(+) w/bump Wing] exhibits the pattern of over-thrombosis. e, H&E images show bleeding in Group 1 as an indication of incomplete closure in contrast to moderate, minimal, and dense thrombotic features observed in Group 2, 3, and 4 respectively as further supported by the signals of activated platelets (green, CD41-positive) and fibrinogen (red) [Scale bars = 0.5 mm (4 mm in box)]. f, Compared to Group 1 [SL(−) HC(−)] and 2 [SL(+) w/flat Wing], Group 3 [SL(+) HC(+)] shows the fastest i) hemostasis and ii) arterial pressure equilibration, indicating the most efficient hemostatic response. g, These outcomes in Group 3 include i) the smallest difference between the proximal and distal pressures with ii) the smallest thrombus area in contrast the largest area of Group 4 [SL(+) w/bump Wing] as an indication of excessive thrombosis. h , The marker gene expression of thrombotic feature (vWF, PF-4, and P-sel) significantly increases from Group 2 to Group 3 and further to Group 4 except the comparison of vWF expression between Group 2 and 3 (ns: no significance). Data are shown as mean ± SD, N = 3 biologically independent animals per group. Significance was determined using one-way ANOVA with Tukey's test between groups.

Article Snippet: Primary antibodies against CD41 (1:100, 24552-1-AP, proteintech), fibrinogen (1:100, ab232793, Abcam), CD31 (1:100, sc-376764, Santa Cruz Biotechnology), CD68 (1:100, ab125212, Abcam), and ARG-1 (1:200, LS-C447907, LSBio) were applied overnight at 4°C.

Techniques: Control, Marker, Gene Expression, Comparison, Expressing

Journal: Clinical and Translational Radiation Oncology

Article Title: Differential regulation of radioadaptation by quercetin between human normal and cancer cells

doi: 10.1016/j.ctro.2025.101099

Figure Lengend Snippet: Quercetin restores NRF2 nuclear translocation in radioadapted MCF10A cells. Representative immunofluorescence images of NRF2 (green) and DAPI (blue) and quantification of the nuclear-to-cytoplasmic NRF2 fluorescence ratio in MCF10A cells 24 h after 5 Gy irradiation with or without prior LDRT and quercetin treatment (Scale bar: 10 μm). Data is shown as mean ± SEM. * p < 0.05, ** p < 0.01.

Article Snippet: Cells were incubated with rabbit anti-human NRF2 primary antibody (1:200; Proteintech, 16396–1-AP), and detection was performed using goat anti-rabbit Alexa Fluor 488 (1:500, Invitrogen, 11001).

Techniques: Translocation Assay, Immunofluorescence, Fluorescence, Irradiation