aggrecan primary antibody  (Proteintech)

Journal: Bioactive Materials

Article Title: Precisely regulated physically-crosslinked carriers enable synergetic release of bioactive factors for MSC-mediated cartilage regeneration

doi: 10.1016/j.bioactmat.2026.01.009

Figure Lengend Snippet: Biocompatibility and Bioactivity of PSF and KSF in vivo . (a) Transwell Assay of MSCs after treated with PBS, MAP and PSF. Scale bar = 200 μm. (b) Wound Healing Assay of MSCs at 0h and 24h. Scale bar = 200 μm. (c) Immunofluorescent staining of cell pellets after 21 days of co-culture. Blue: DAPI; Green: ActinGreen; Red: Aggrecan. Scale bar = 200 μm. (d) Alcian blue staining of 2D cultured MSCs after 14 days. Scale bar = 200 μm. (e) Cell viability of MSCs at day 3 after co-culture. (f) The cell number of MSCs migrated from upper to lower chamber in Transwell assay. (g) The average distance MSCs migrated from injured margin in wound healing assay. (h–j) qRT-PCR of Col2a1 , Acan and Sox9 mRNA relative expression ratio compared with PBS group. ns: p > 0.05; ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001.

Article Snippet: Immunofluorescent staining was performed with Aggrecan primary antibody (13880-1-AP, Proteintech, USA), ActinGreen ( R37110 , Thermo, USA) and DAPI (Solarbio, China) and observed with 3D reconstruction under high content imaging system (PerkinElmer, Operetta CLS, USA).

Techniques: In Vivo, Transwell Assay, Wound Healing Assay, Staining, Co-Culture Assay, Cell Culture, Quantitative RT-PCR, Expressing

Journal: Nature Communications

Article Title: Targeting the ROS-ferroptosis-inflammation cycle with a nanozyme-functionalized hydrogel for intervertebral disc repair

doi: 10.1038/s41467-025-66116-w

Figure Lengend Snippet: a, b Live/dead staining ( a ) and its quantification ( b ) of TBHP-pretreated NPCs with or without hydrogel co-culture. Scale bar: 50 μm. c, d Intracellular ROS levels detected by DCFH-DA staining ( c ) and its quantification ( d ). Scale bar: 50 μm. e, f Representative flow cytometry plots ( e ) and quantification ( f ) of intracellular ROS levels. MFI, mean fluorescence intensity. g, h Representative flow cytometry plots ( g ) and quantification ( h ) of mitochondrial ROS levels. i, j Lipid peroxidation (LPO) staining ( i ) and its quantification ( j ) in TBHP-pretreated NPCs co-cultured with or without hydrogels. Scale bar: 10 μm. k, l FerroOrange staining ( k ) and its quantification ( l ). Scale bar: 20 μm. m, n Representative flow cytometry plots ( m ) and statistical analysis ( n ) of lipid peroxidation levels. o, p Representative flow cytometry plots ( o ) and statistical analysis ( p ) of ferrous ion levels. q, r GPX4 immunofluorescence ( q ) and its quantification ( r ) in TBHP-pretreated NPCs co-cultured with or without hydrogels. Scale bar: 20 μm. s, t Aggrecan (ACAN) immunofluorescence ( s ) and its quantification ( t ). Scale bar: 20 μm. u, v Representative western blots ( u ) of ACSL4, GPX4, MMP3, and MMP13 in TBHP-pretreated NPCs co-cultured with or without hydrogels and corresponding statistical analyses ( v ). The untreated NPCs served as the control (Con) group. Data are presented as mean ± SD from three independent replicates, and p -values were calculated using one-way ANOVA with Tukey’s multiple comparisons test. Source data are provided as a file.

Article Snippet: Immunohistochemical staining was conducted on these sections using primary antibodies against ACAN (Proteintech, 13880-1-AP), FTH1 (ABclonal, A19544), ACSL4 (Santa Cruz, sc-365230), and GPX4 (Proteintech, 67763-1-Ig) (all at 1:100 dilution), followed by overnight incubation at 4 °C.

Techniques: Staining, Co-Culture Assay, Flow Cytometry, Fluorescence, Cell Culture, Immunofluorescence, Western Blot, Control

Journal: Nature Communications

Article Title: Targeting the ROS-ferroptosis-inflammation cycle with a nanozyme-functionalized hydrogel for intervertebral disc repair

doi: 10.1038/s41467-025-66116-w

Figure Lengend Snippet: a Schematic illustration of in vivo animal experiment design was created in BioRender (Wang, Y. 2025, https://BioRender.com/bqmgjz2 ). b, c X-ray ( b ) and MRI ( c ) images of rat caudal disks from the control (Con), degenerated (IDD), and PG@MBC-treated (PG@MBC) groups. d Quantification of the disc height index (DHI). e Relative water content of rat NP tissues based on MRI results. f Quantification of the degenerative grades of rat disks. g, h SO/FG staining images of rat disks ( h ) and the corresponding histological grades ( g ). Scale bar: 1 mm. i, j Immunohistochemical staining of aggrecan (ACAN) ( i ) and corresponding quantification ( j ). Scale bar: 1 mm. k, l Immunohistochemical staining of GPX4 ( k ) and corresponding quantification ( l ). Scale bar: 1 mm. m, n Quantification of malondialdehyde (MDA) ( m ) and glutathione (GSH) ( n ) concentrations in rat NP tissues. o Temporal changes in Il6 mRNA levels in rat NP tissues. p Temporal changes in IL6 secretion in rat NP tissues. q, r Immunofluorescence staining ( q ) of HuR and p-STAT3 in rat NP tissues at 3 weeks post-surgery and corresponding quantification of mean fluorescence intensity (MFI) ( r ). s-u Compression curves ( s ), compression modulus ( t ), and compression cycle curves ( u ) of rat disks at 6 weeks post-surgery. v-x Compression curves ( v ), compression modulus ( w ), and compression cycle curves ( x ) of rat disks at 12 weeks post-surgery. n = 5 independent replicates for panels d-g . n = 3 independent replicates for panels j, l–p, r, t , w . Data are presented as mean ± SD, and p -values were calculated using one-way ANOVA with Tukey’s multiple comparisons test ( d, e, j, l–p, r, t , w ) and Kruskal-Wallis test ( f, g ). Source data are provided as a file.

Article Snippet: Immunohistochemical staining was conducted on these sections using primary antibodies against ACAN (Proteintech, 13880-1-AP), FTH1 (ABclonal, A19544), ACSL4 (Santa Cruz, sc-365230), and GPX4 (Proteintech, 67763-1-Ig) (all at 1:100 dilution), followed by overnight incubation at 4 °C.

Techniques: In Vivo, Control, Staining, Immunohistochemical staining, Immunofluorescence, Fluorescence

Journal: Journal of Orthopaedic Translation

Article Title: Establishment and characterization of an inflammatory cartilaginous organoids model for organoid transplantation study

doi: 10.1016/j.jot.2025.05.002

Figure Lengend Snippet: Characterization of cartilaginous and inflammatory organoids. (a) The Alcian blue staining was performed on cartilaginous organoids of day 28. Scale bar = 100 μm. (b) The Safranin O and Fast Green staining was performed on the cartilaginous organoids of day 28. The Immunofluorescence staining of the (c) DAPI, (d) ACAN and (e) merged image of cartilaginous organoids. The Immunofluorescence staining of the (f) DAPI, (g) COL2A1 and (h) merged image of cartilaginous organoids. Scale bar = 100 μm. N = 3.

Article Snippet: 10 μL of the anti-COL2A1 primary antibody (Proteintech, #28459-1-AP) and the anti-ACAN primary antibody (Proteintech, #13880-1-AP) were dripped and incubated at 4 °C overnight respectively.

Techniques: Staining, Immunofluorescence