Anti-ADAMTS5 Antibody Picoband®

Western blot analysis of ADAMTS5 using anti-ADAMTS5 antibody (A02802-1).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human Hela whole cell lysates,
Lane 2: human HepG2 whole cell lysates,
Lane 3: human U20S whole cell lysates,
Lane 4: human SW620 whole cell lysates,
Lane 5: human THP-1 whole cell lysates,
Lane 6: human Raji whole cell lysates,
Lane 7: human SGC-7901 whole cell lysates,
Lane 8: rat liver tissue lysates,
Lane 9: rat testis tissue lysates,
Lane 10: mouse liver tissue lysates,
Lane 11: mouse testis tissue lysates,
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-ADAMTS5 antigen affinity purified polyclonal antibody (Catalog # A02802-1) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for ADAMTS5 at approximately 75 kDa. The expected band size for ADAMTS5 is at 102 kDa.

PA suppressed excess expression of the catabolic indicators of chondrocytes induced by IL-1β, including ADAMTS5, MMP1, MMP3, and MMP13. Mice chondrocytes were treated with 5 ng/ml of IL-1β, alone or with PA (2.5, 5, and 10 μM) for 24 h (A) Western blotting results and (B–E) quantitative analysis of ADAMTS5, MMP1, MMP3, and MMP13. (F) MMP13 expression was observed by immunofluorescence staining when chondrocytes were treated with 5 ng/ml of IL-1β, alone or with 10 μM of PA (scale bar 200 μm). (G–I) Relative mRNA levels of ADAMTS5, MMP3, and MMP13 in chondrocytes stimulated with 5 ng/ml of IL-1β, alone or with PA (2.5, 5, and 10 μM) for 24 h. GAPDH was used as an internal reference. Data are presented as means ± SD ( n = 3). The exact p value was marked in the corresponding figure and p < 0.05 was considered statistically significant.
Index in PubMed under a CC BY license. PMID: 34925020

Knockdown of integrin αVβ3 weakened the anti-inflammatory, anabolism enhancing, and catabolism inhibiting effect of PA on IL-1β-induced chondrocytes. (A,B) Relative mRNA levels of integrin αV (Itg αV) and integrin β3 (Itg β3) in chondrocytes stimulated with 5 ng/ml of IL-1β, alone or with PA (2.5, 5, and 10 μM) for 24 h (C,D) Itg αV and Itg β3 were knocked down by siRNA transfection, and the knockdown efficiency was verified by RT-PCR. (E,F) Inflammatory markers (COX2, iNOS) were detected by western blotting and the band density of protein levels were quantified after mice chondrocytes were added with or without 5 ng/ml of IL-1β, 10 μM of PA, and Itg αVβ3 siRNA. (G–I) Western blotting was applied to measure the anabolic (aggrecan, collagen II) and catabolic markers (MMP1, MMP3, MMP13, and ADAMTS5) in the Itg αVβ3-deficiency mice chondrocytes along with or without the administration of 5 ng/ml of IL-1β and 10 μM of PA, and the band density of these protein levels were quantified in the histogram. GAPDH was used as an internal reference. Data are presented as means ± SD ( n = 3). The exact p value was marked in the corresponding figure and p < 0.05 was considered statistically significant.
Index in PubMed under a CC BY license. PMID: 34925020

ALT alleviated excess MMPs and ADAMTS5 expression in IL-1β-stimulated mouse chondrocytes. Chondrocytes were exposed to ALT (2.5, 5, and 10 μM) with or without IL-1β (10 ng/ml) for 24 h. (A) Western blot was employed to determine the expression of MMP1, MMP3, MMP13, and ADAMTS5. (B) Relative protein expression was qualified by ImageJ software, GAPDH was used as the internal control ( n = 3). # p < 0.05 vs. control group; * p < 0.05 vs. IL-1β group; ** p < 0.01 vs. IL-1β group.
Index in PubMed under a CC BY license. PMID: 34650433

Schematic diagram of the effect of ALT on cartilage degeneration. IL-1β induces the expression of pro-inflammatory factors, including iNOS, COX2, MMPs and ADAMTS5. IL-1β stimulates the degradation of Collagen II and the impairment of autophagy. Furthermore, IL-1β functions by activating the PI3K/AKT/mTOR, NF-κB signaling pathways, phosphorylating and translocating STAT3. As shown in the figure, the levels of P-PI3K, P-AKT, P-mTOR, P-IKB, P-P65, P-STAT3 and the translocation of P65 and STAT3 are increasing. However, ALT can attenuate these effects.
Index in PubMed under a CC BY license. PMID: 34650433

Effects of Schisandrin A on IL-1β-induced MMPs and ADAMTS5 protein expression. Chondrocytes were exposed to Schisandrin A (25, 50 μM) with or without IL-1β (10 ng/ml) for 24 h. (A) Western blot was employed to determine the expression of MMP1, MMP3, MMP13, and ADAMTS5. (B) Relative protein expression was qualified by Image-J software, GAPDH was used as the internal control ( n = 3). # P < 0.05 vs. control group; ∗ P < 0.05 vs. IL-1β group; ∗∗ P < 0.01 vs. IL-1β group.
Index in PubMed under a CC BY license. PMID: 30761007

IHC analysis of ADAMTS5 using anti-ADAMTS5 antibody (A02802-1).
ADAMTS5 was detected in a paraffin-embedded section of human placenta tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-ADAMTS5 Antibody (A02802-1) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.