Incubation:Article Title: Metformin Ameliorates Senescence of Adipose-Derived Mesenchymal Stem Cells and Attenuates Osteoarthritis Progression via the AMPK-Dependent Autophagy Pathway
Article Snippet: .. The sections were separately incubated with mouse anti-COL2A1 (Santa Cruz Biotechnology, Texas, USA, 1 : 50), rabbit anti-MMP-13 (Bioss, Beijing, China, 1 : 200), rabbit anti-F4/80 (Cell Signaling, 1: 400), rabbit anti-iNOS (Abcam, 1 : 100), and rabbit anti-CD206 (Abcam, 1 : 1000) overnight at 4°C. .. Then, the slides were incubated with a biotin-labeled IgG secondary antibody (Zhongshan Golden Bridge, Beijing, China) and stained with a diaminobenzidine (DAB) substrate and counterstained with hematoxylin.
Article Title: Surface-Engineered Umbilical Cord Mesenchymal Stem Cell-Derived sEVs for Targeted Therapy of Osteoarthritis.
Article Snippet: Small extracellular vesicles (sEVs) from human umbilical-cord-derived mesenchymal stem cells (UCMSCs) hold promise for cartilage regeneration in osteoarthritis (OA).. However, their therapeutic effectiveness is significantly limited by rapid clearance through blood and lymphatic vessels in the synovial tissue after intra-articular injection.. This study aimed to enhance the chondrocyte-targeting ability and improve the cartilage repair potential of UCMSC-derived sEVs by conjugating them with a chondrocyte-affinity peptide (CAP) using copper-free click chemistry.
Article Title: A Highly Conserved Shh Enhancer Coordinates Hypothalamic and Craniofacial Development
Article Snippet: .. Slides were incubated in primary antibodies diluted in blocking solution overnight at 4°C as follows: rabbit anti-CASP3 (1:500, Abcam, ab49822); mouse anti-COL2A1 (1:400, Santa Cruz, sc-52659); rabbit anti-ISL1 (1:200, Abcam, ab20670); rabbit anti-LHX3 (1:250, Abcam, ab14555); rabbit anti-NKX2-1 (1:250, Abcam, ab76013); rabbit anti-NKX2-2 (1:250, Abcam, ab191077); rabbit anti-NKX6-1 (1:250, Abcam, ab221549); rabbit anti-OLIG2 (1:200, Abcam, ab109186); rabbit anti-oxytocin (1:500, Abcam, ab212193); rabbit anti-PAX6 (1:350, Abcam, ab195045); sheep anti-PITX2 (1:200, R&D, AF7388); rabbit anti-SF1 (1:200, Abcam, ab65815); rabbit anti-TBX3 (1:200, Abcam, ab99302); rabbit anti-TH1 (1:200, Abcam, ab137869); rabbit anti-TCF4 (1:100, Cell Signalling; C48H11); rabbit anti-vasopressin (1:500, Abcam, ab213707). .. Slides were then washed 3 times for 10 min each in PBT and incubated for 1 h at room temperature with fluorescent secondary antibodies in PBT in the dark, secondaries as follows: goat anti-rabbit 488 (1:500, Thermo Fisher Scientific, A11034); goat anti-mouse 488 (1:500, Thermo Fisher Scientific, A11029); donkey anti-sheep 594 (1:500, Thermo Fisher Scientific, A11016).
Article Title: A Highly Conserved Shh Enhancer Coordinates Hypothalamic and Craniofacial Development
Article Snippet: .. For PITX2 staining, donkey serum was used, Slides were incubated in primary antibodies diluted in blocking solution overnight at 4°C as follows: rabbit anti-CASP3 (1:500, Abcam, ab49822); mouse anti-COL2A1 (1:400, Santa Cruz, sc-52659); rabbit anti-ISL1 (1:200, Abcam, ab20670); rabbit anti-LHX3 (1:250, Abcam, ab14555); rabbit anti-NKX2-1 (1:250, Abcam, ab76013); rabbit anti-NKX2-2 (1:250, Abcam, ab191077); rabbit anti-NKX6-1 (1:250, Abcam, ab221549); rabbit anti-OLIG2 (1:200, Abcam, ab109186); rabbit anti-oxytocin (1:500, Abcam, ab212193); rabbit anti-PAX6 (1:350, Abcam, ab195045); sheep anti-PITX2 (1:200, R&D, AF7388); rabbit anti-SF1 (1:200, Abcam, ab65815); rabbit anti-TBX3 (1:200, Abcam, ab99302); rabbit anti-TH1 (1:200, Abcam, ab137869); rabbit anti-TCF4 (1:100, Cell Signalling; C48H11); rabbit anti-vasopressin (1:500, Abcam, ab213707). .. Slides were then washed 3 times for 10 min each in PBT and incubated for 1 h at room temperature with fluorescent secondary antibodies in PBT in the dark, secondaries as follows: goat anti-rabbit 488 (1:500, Thermo Fisher, A11034); goat anti-mouse 488 (1:500, Thermo Fisher, A11029); donkey anti-sheep 594 (1:500, Thermo Fisher, A11016).
Blocking Assay:Article Title: Surface-Engineered Umbilical Cord Mesenchymal Stem Cell-Derived sEVs for Targeted Therapy of Osteoarthritis.
Article Snippet: Small extracellular vesicles (sEVs) from human umbilical-cord-derived mesenchymal stem cells (UCMSCs) hold promise for cartilage regeneration in osteoarthritis (OA).. However, their therapeutic effectiveness is significantly limited by rapid clearance through blood and lymphatic vessels in the synovial tissue after intra-articular injection.. This study aimed to enhance the chondrocyte-targeting ability and improve the cartilage repair potential of UCMSC-derived sEVs by conjugating them with a chondrocyte-affinity peptide (CAP) using copper-free click chemistry.
Article Title: A Highly Conserved Shh Enhancer Coordinates Hypothalamic and Craniofacial Development
Article Snippet: .. Slides were incubated in primary antibodies diluted in blocking solution overnight at 4°C as follows: rabbit anti-CASP3 (1:500, Abcam, ab49822); mouse anti-COL2A1 (1:400, Santa Cruz, sc-52659); rabbit anti-ISL1 (1:200, Abcam, ab20670); rabbit anti-LHX3 (1:250, Abcam, ab14555); rabbit anti-NKX2-1 (1:250, Abcam, ab76013); rabbit anti-NKX2-2 (1:250, Abcam, ab191077); rabbit anti-NKX6-1 (1:250, Abcam, ab221549); rabbit anti-OLIG2 (1:200, Abcam, ab109186); rabbit anti-oxytocin (1:500, Abcam, ab212193); rabbit anti-PAX6 (1:350, Abcam, ab195045); sheep anti-PITX2 (1:200, R&D, AF7388); rabbit anti-SF1 (1:200, Abcam, ab65815); rabbit anti-TBX3 (1:200, Abcam, ab99302); rabbit anti-TH1 (1:200, Abcam, ab137869); rabbit anti-TCF4 (1:100, Cell Signalling; C48H11); rabbit anti-vasopressin (1:500, Abcam, ab213707). .. Slides were then washed 3 times for 10 min each in PBT and incubated for 1 h at room temperature with fluorescent secondary antibodies in PBT in the dark, secondaries as follows: goat anti-rabbit 488 (1:500, Thermo Fisher Scientific, A11034); goat anti-mouse 488 (1:500, Thermo Fisher Scientific, A11029); donkey anti-sheep 594 (1:500, Thermo Fisher Scientific, A11016).
Article Title: A Highly Conserved Shh Enhancer Coordinates Hypothalamic and Craniofacial Development
Article Snippet: .. For PITX2 staining, donkey serum was used, Slides were incubated in primary antibodies diluted in blocking solution overnight at 4°C as follows: rabbit anti-CASP3 (1:500, Abcam, ab49822); mouse anti-COL2A1 (1:400, Santa Cruz, sc-52659); rabbit anti-ISL1 (1:200, Abcam, ab20670); rabbit anti-LHX3 (1:250, Abcam, ab14555); rabbit anti-NKX2-1 (1:250, Abcam, ab76013); rabbit anti-NKX2-2 (1:250, Abcam, ab191077); rabbit anti-NKX6-1 (1:250, Abcam, ab221549); rabbit anti-OLIG2 (1:200, Abcam, ab109186); rabbit anti-oxytocin (1:500, Abcam, ab212193); rabbit anti-PAX6 (1:350, Abcam, ab195045); sheep anti-PITX2 (1:200, R&D, AF7388); rabbit anti-SF1 (1:200, Abcam, ab65815); rabbit anti-TBX3 (1:200, Abcam, ab99302); rabbit anti-TH1 (1:200, Abcam, ab137869); rabbit anti-TCF4 (1:100, Cell Signalling; C48H11); rabbit anti-vasopressin (1:500, Abcam, ab213707). .. Slides were then washed 3 times for 10 min each in PBT and incubated for 1 h at room temperature with fluorescent secondary antibodies in PBT in the dark, secondaries as follows: goat anti-rabbit 488 (1:500, Thermo Fisher, A11034); goat anti-mouse 488 (1:500, Thermo Fisher, A11029); donkey anti-sheep 594 (1:500, Thermo Fisher, A11016).
other:
Staining:Article Title: A Highly Conserved Shh Enhancer Coordinates Hypothalamic and Craniofacial Development
Article Snippet: .. For PITX2 staining, donkey serum was used, Slides were incubated in primary antibodies diluted in blocking solution overnight at 4°C as follows: rabbit anti-CASP3 (1:500, Abcam, ab49822); mouse anti-COL2A1 (1:400, Santa Cruz, sc-52659); rabbit anti-ISL1 (1:200, Abcam, ab20670); rabbit anti-LHX3 (1:250, Abcam, ab14555); rabbit anti-NKX2-1 (1:250, Abcam, ab76013); rabbit anti-NKX2-2 (1:250, Abcam, ab191077); rabbit anti-NKX6-1 (1:250, Abcam, ab221549); rabbit anti-OLIG2 (1:200, Abcam, ab109186); rabbit anti-oxytocin (1:500, Abcam, ab212193); rabbit anti-PAX6 (1:350, Abcam, ab195045); sheep anti-PITX2 (1:200, R&D, AF7388); rabbit anti-SF1 (1:200, Abcam, ab65815); rabbit anti-TBX3 (1:200, Abcam, ab99302); rabbit anti-TH1 (1:200, Abcam, ab137869); rabbit anti-TCF4 (1:100, Cell Signalling; C48H11); rabbit anti-vasopressin (1:500, Abcam, ab213707). .. Slides were then washed 3 times for 10 min each in PBT and incubated for 1 h at room temperature with fluorescent secondary antibodies in PBT in the dark, secondaries as follows: goat anti-rabbit 488 (1:500, Thermo Fisher, A11034); goat anti-mouse 488 (1:500, Thermo Fisher, A11029); donkey anti-sheep 594 (1:500, Thermo Fisher, A11016).
|