ADAMTS5 Antibody - #DF13268

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产品: ADAMTS5 抗体
货号: DF13268
描述: Rabbit polyclonal antibody to ADAMTS5
应用: WB IHC
反应: Human, Mouse, Rat
预测: Pig, Bovine, Horse, Sheep, Rabbit, Dog, Chicken, Xenopus
分子量: 72 kd; 102kD(Calculated).
蛋白号: Q9UNA0
RRID: AB_2846287

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产品描述

来源:
Rabbit
应用:
WB 1:500-1:2000, IHC 1:50-1:200
*The optimal dilutions should be determined by the end user.
*Tips:

WB: 适用于变性蛋白样本的免疫印迹检测. IHC: 适用于组织样本的石蜡(IHC-p)或冰冻(IHC-f)切片样本的免疫组化/荧光检测. IF/ICC: 适用于细胞样本的荧光检测. ELISA(peptide): 适用于抗原肽的ELISA检测.

反应:
Human,Mouse,Rat
预测:
Pig(100%), Bovine(100%), Horse(100%), Sheep(100%), Rabbit(100%), Dog(100%), Chicken(100%), Xenopus(100%)
克隆:
Polyclonal
特异性:
ADAMTS5 Antibody detects endogenous levels of total ADAMTS5.
RRID:
AB_2846287
引用格式: Affinity Biosciences Cat# DF13268, RRID:AB_2846287.
偶联:
Unconjugated.
纯化:
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
保存:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
别名:

展开/折叠

A disintegrin and metalloproteinase with thrombospondin motifs 11; A disintegrin and metalloproteinase with thrombospondin motifs 5; A disintegrin like and metalloprotease (reprolysin type) with thrombospondin type 1 motif 5; A disintegrin like and metalloprotease (reprolysin type) with thrombospondin type 1 motif, 5 (aggrecanase 2); A disintegrin-like and metalloprotease with thrombospondin type 1 motif, 5; A Disintigrin And Metalloproteinase with ThromboSpondin motif-5; ADAM metallopeptidase with thrombospondin type 1 motif 5; ADAM TS 11; ADAM TS 5; ADAM TS5; ADAM-TS 11; ADAM-TS 5; ADAM-TS5; ADAMTS 11; ADAMTS 5; ADAMTS-11; ADAMTS-5; ADAMTS11; ADAMTS11, formerly; Adamts5; ADMP 2; ADMP-2; ADMP2; Aggrecanase 2; Aggrecanase-2; ATS5_HUMAN; FLJ36738; Implantin; ThromboSpondin motif-5;

抗原和靶标

免疫原:
Uniprot:

Q9UNA0(ATS5_HUMAN) >>Visit HPA database.

基因/基因ID:
ADAMTS5(11096)
表达:
Q9UNA0 ATS5_HUMAN:

Expressed at low level in placenta primarily but also detected in heart and brain, cervix, uterus, bladder, esophagus, rib cartilage, chondroblastoma, fibrous tissue and a joint capsule from an arthritic patient.

序列:
MLLGWASLLLCAFRLPLAAVGPAATPAQDKAGQPPTAAAAAQPRRRQGEEVQERAEPPGHPHPLAQRRRSKGLVQNIDQLYSGGGKVGYLVYAGGRRFLLDLERDGSVGIAGFVPAGGGTSAPWRHRSHCFYRGTVDGSPRSLAVFDLCGGLDGFFAVKHARYTLKPLLRGPWAEEEKGRVYGDGSARILHVYTREGFSFEALPPRASCETPASTPEAHEHAPAHSNPSGRAALASQLLDQSALSPAGGSGPQTWWRRRRRSISRARQVELLLVADASMARLYGRGLQHYLLTLASIANRLYSHASIENHIRLAVVKVVVLGDKDKSLEVSKNAATTLKNFCKWQHQHNQLGDDHEEHYDAAILFTREDLCGHHSCDTLGMADVGTICSPERSCAVIEDDGLHAAFTVAHEIGHLLGLSHDDSKFCEETFGSTEDKRLMSSILTSIDASKPWSKCTSATITEFLDDGHGNCLLDLPRKQILGPEELPGQTYDATQQCNLTFGPEYSVCPGMDVCARLWCAVVRQGQMVCLTKKLPAVEGTPCGKGRICLQGKCVDKTKKKYYSTSSHGNWGSWGSWGQCSRSCGGGVQFAYRHCNNPAPRNNGRYCTGKRAIYRSCSLMPCPPNGKSFRHEQCEAKNGYQSDAKGVKTFVEWVPKYAGVLPADVCKLTCRAKGTGYYVVFSPKVTDGTECRLYSNSVCVRGKCVRTGCDGIIGSKLQYDKCGVCGGDNSSCTKIVGTFNKKSKGYTDVVRIPEGATHIKVRQFKAKDQTRFTAYLALKKKNGEYLINGKYMISTSETIIDINGTVMNYSGWSHRDDFLHGMGYSATKEILIVQILATDPTKPLDVRYSFFVPKKSTPKVNSVTSHGSNKVGSHTSQPQWVTGPWLACSRTCDTGWHTRTVQCQDGNRKLAKGCPLSQRPSAFKQCLLKKC

种属预测

种属预测:

score>80的预测可信度较高，可尝试用于WB检测。*预测模型主要基于免疫原序列比对，结果仅作参考，不作为质保凭据。

Species
Results
Score
Pig
100
Horse
100
Bovine
100
Sheep
100
Dog
100
Xenopus
100
Chicken
100
Rabbit
100
Zebrafish
67
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

翻译修饰 - Q9UNA0 作为底物

Site PTM Type Enzyme
T36 O-Glycosylation
Y81 Phosphorylation
Y89 Phosphorylation
Y92 Phosphorylation
S208 O-Glycosylation
S214 O-Glycosylation
T215 O-Glycosylation
S440 Phosphorylation
S441 Phosphorylation
T444 Phosphorylation
N498 N-Glycosylation
W570 C-Glycosylation
W573 C-Glycosylation
S582 O-Glycosylation
K720 Ubiquitination
T746 Phosphorylation
K858 Acetylation
T899 Phosphorylation

研究背景

功能:

Metalloproteinase that plays an important role in connective tissue organization, development, inflammation, arthritis, and cell migration. ADAMTS5 is an extracellular matrix (ECM) degrading enzyme that show proteolytic activity toward the hyalectan group of chondroitin sulfate proteoglycans (CSPGs) including aggrecan, versican, brevican and neurocan. Cleavage within the hyalectans occurs at Glu-Xaa recognition motifs. Plays a role in embryonic development, including limb and cardiac morphogenesis, and skeletal muscle development through its versican remodeling properties. Participates in development of brown adipose tissue and browning of white adipose tissue. Plays an important role for T-lymphocyte migration from draining lymph nodes following viral infection.

翻译修饰:

The precursor is cleaved by furin and PCSK7 outside of the cell.

C- and O-glycosylated. O-fucosylated by POFUT2 on a serine or a threonine residue found within the consensus sequence C1-X(2)-(S/T)-C2-G of the TSP type-1 repeat domains where C1 and C2 are the first and second cysteine residue of the repeat, respectively. Fucosylated repeats can then be further glycosylated by the addition of a beta-1,3-glucose residue by the glucosyltransferase, B3GALTL. Fucosylation can mediate the efficient secretion of ADAMTS family members. Can be C-glycosylated with one or two mannose molecules on tryptophan residues within the consensus sequence W-X-X-W of the TPRs, and N-glycosylated. These other glycosylations can also facilitate secretion (By similarity).

Glycosylated. Can be O-fucosylated by POFUT2 on a serine or a threonine residue found within the consensus sequence C1-X(2)-(S/T)-C2-G of the TSP type-1 repeat domains where C1 and C2 are the first and second cysteine residue of the repeat, respectively. Fucosylated repeats can then be further glycosylated by the addition of a beta-1,3-glucose residue by the glucosyltransferase, B3GALTL. Fucosylation mediates the efficient secretion of ADAMTS family members. Also can be C-glycosylated with one or two mannose molecules on tryptophan residues within the consensus sequence W-X-X-W of the TPRs, and N-glycosylated. These other glycosylations can also facilitate secretion (By similarity).

细胞定位:

Secreted>Extracellular space>Extracellular matrix.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
组织特异性:

Expressed at low level in placenta primarily but also detected in heart and brain, cervix, uterus, bladder, esophagus, rib cartilage, chondroblastoma, fibrous tissue and a joint capsule from an arthritic patient.

蛋白家族:

The spacer domain and the TSP type-1 domains are important for a tight interaction with the extracellular matrix.

The conserved cysteine present in the cysteine-switch motif binds the catalytic zinc ion, thus inhibiting the enzyme. The dissociation of the cysteine from the zinc ion upon the activation-peptide release activates the enzyme.

文献引用

1). Dual functions of microRNA-17 in maintaining cartilage homeostasis and protection against osteoarthritis. Nature Communications, 2022 (PubMed: 35508470) [IF=16.6]

Application: WB    Species: Mouse    Sample:

Fig. 2 miR-17 inhibits cartilage destruction by targeting multiple pathological catabolic factors. a Luciferase activity of 293 T cells co-transfected with dual-luciferase reporter constructs containing wild-type or mutated 3′-UTRs, as well as negative control (mimic NC) or miR-17 mimic. The data presented are mean percentage changes over mimic NC ± s.e.m. n = 6 biologically independent samples (NC + wt UTR, miR-17+wt UTR); n = 4 biologically independent samples (NC + mut UTR, miR-17 + mut UTR). b Mouse articular chondrocytes were transfected with miR-17 mimic (50 nM) or mimic NC and treated with or without IL-1β (5 ng/mL) for 24 h. The protein levels of catabolic and anabolic factors were determined by western blot followed by densitometry analysis. Blots are representative of three independent experiments. c, d qRT-PCR analysis of catabolic genes in knee cartilage (c), and representative images of IHC staining and quantification of MMP13+, ADAMTS5+ and NOS2+ cells in joint sections (d) of mice subjected to sham or DMM surgery and intra-articular injections of agomir-NC or agomir-17 (1.5 nmol) for 4 weeks, beginning at 4 weeks after surgery. n = 5 biologically independent samples per group in c. For MMP13 and ADAMTS5 staining, n = 4 mice (sham); n = 5 mice (DMM, DMM + agomir-17). For NOS2 staining, n = 4 mice per group. e, f qRT-PCR analysis of catabolic genes (e) and representative images of MMP13, ADAMTS5, and NOS2 immunostaining, and quantification of positive cells (f) from knee joints of cKO mice subjected to DMM surgery and intra-articular injection of agomir-NC, or agomir-17 (2 nmol). The injections were performed at 1 week after surgery and samples were collected at 2.5 weeks after surgery. n = 4 biologically independent samples per group in (e); n = 4 mice per group in (f). All scale bars, 100 μm. Data are presented as mean ± s.e.m. or boxplots (center line, median; box limits, 25 to 75th percentiles; whiskers, min to max), and dots represent individual mice or biologically independent samples. NS, not significant, *P < 0.05, **P < 0.01, and ***P < 0.001 in (a, c–f). *P < 0.05 versus control group, °P < 0.05 versus IL-1β + mimic NC in (b). Two-sided Student’s t test for (a); one-way ANOVA with Bonferroni’s test for (b–f). Exact P values in (b): *P = 0.0011, °P = 0.0205 (MMP3); *P <0.0001, °P = 0.0039 (MMP13); *P = 0.0015, °P = 0.0040 (ADAMTS5); *P = 0.0001, °P = 0.0013 (NOS2); *P = 0.0013, P > 0.9999 (SOX9); *P = 0.0001, P > 0.9999 (COL2A1). Source data are provided as a Source Data file.

Application: IHC    Species: Mouse    Sample:

Fig. 2 miR-17 inhibits cartilage destruction by targeting multiple pathological catabolic factors. a Luciferase activity of 293 T cells co-transfected with dual-luciferase reporter constructs containing wild-type or mutated 3′-UTRs, as well as negative control (mimic NC) or miR-17 mimic. The data presented are mean percentage changes over mimic NC ± s.e.m. n = 6 biologically independent samples (NC + wt UTR, miR-17+wt UTR); n = 4 biologically independent samples (NC + mut UTR, miR-17 + mut UTR). b Mouse articular chondrocytes were transfected with miR-17 mimic (50 nM) or mimic NC and treated with or without IL-1β (5 ng/mL) for 24 h. The protein levels of catabolic and anabolic factors were determined by western blot followed by densitometry analysis. Blots are representative of three independent experiments. c, d qRT-PCR analysis of catabolic genes in knee cartilage (c), and representative images of IHC staining and quantification of MMP13+, ADAMTS5+ and NOS2+ cells in joint sections (d) of mice subjected to sham or DMM surgery and intra-articular injections of agomir-NC or agomir-17 (1.5 nmol) for 4 weeks, beginning at 4 weeks after surgery. n = 5 biologically independent samples per group in c. For MMP13 and ADAMTS5 staining, n = 4 mice (sham); n = 5 mice (DMM, DMM + agomir-17). For NOS2 staining, n = 4 mice per group. e, f qRT-PCR analysis of catabolic genes (e) and representative images of MMP13, ADAMTS5, and NOS2 immunostaining, and quantification of positive cells (f) from knee joints of cKO mice subjected to DMM surgery and intra-articular injection of agomir-NC, or agomir-17 (2 nmol). The injections were performed at 1 week after surgery and samples were collected at 2.5 weeks after surgery. n = 4 biologically independent samples per group in (e); n = 4 mice per group in (f). All scale bars, 100 μm. Data are presented as mean ± s.e.m. or boxplots (center line, median; box limits, 25 to 75th percentiles; whiskers, min to max), and dots represent individual mice or biologically independent samples. NS, not significant, *P < 0.05, **P < 0.01, and ***P < 0.001 in (a, c–f). *P < 0.05 versus control group, °P < 0.05 versus IL-1β + mimic NC in (b). Two-sided Student’s t test for (a); one-way ANOVA with Bonferroni’s test for (b–f). Exact P values in (b): *P = 0.0011, °P = 0.0205 (MMP3); *P <0.0001, °P = 0.0039 (MMP13); *P = 0.0015, °P = 0.0040 (ADAMTS5); *P = 0.0001, °P = 0.0013 (NOS2); *P = 0.0013, P > 0.9999 (SOX9); *P = 0.0001, P > 0.9999 (COL2A1). Source data are provided as a Source Data file.
2). Sustained therapeutic effects of self-assembled hyaluronic acid nanoparticles loaded with α-Ketoglutarate in various osteoarthritis stages. Biomaterials, 2024 (PubMed: 39326362) [IF=12.8]
3). Pentraxin 3 regulated by miR-224-5p modulates macrophage reprogramming and exacerbates osteoarthritis associated synovitis by targeting CD32. Cell Death & Disease, 2022 (PubMed: 35739102) [IF=8.1]

Application: IF/ICC    Species: Mouse    Sample:

Fig. 2 PTX3 accelerates synovitis, cartilage erosion, and exacerbates OA development in mice. A, B Safranin O/fast green staining and Osteoarthritis Research Society International (OARSI) grades of knee cartilage from DMM mice treated with vehicle, recombinant mouse PTX3 (rmPTX3), or PTX3 neutralizing antibody (PTX3-NAb) for 4 weeks and 8 weeks. Scale bar: 100 µm. C–F IF staining and quantification of ADATMS5 and MMP13 in knee cartilage of controls and DMM mice treated with vehicle, rmPTX3, or PTX3-NAb for 8 weeks. Scale bar: 50 µm. G IF staining and quantification of COL2 in knee cartilage of controls and DMM mice treated with rmPTX3 or PTX3-NAb for 8 weeks. Scale bar: 50 µm. H, I H&E and synovitis score of synovial tissues from DMM mice treated with vehicle, rmPTX3, or PTX3-NAb for 4 weeks and 8 weeks. Scale bar: 100 µm. *P 
4). TMF inhibits extracellular matrix degradation by regulating the C/EBPβ/ADAMTS5 signaling pathway in osteoarthritis. Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie, 2024 (PubMed: 38554527) [IF=6.9]

Application: WB    Species: Human    Sample: C28/I2 cells

Fig. 4. Knockdown of FOXO3a compromised the inhibitory effects of TMF on C/EBPβ-mediated ECM degradation. (A–B) IHC assays were used to detect the altered expression of FOXO3a. (C–D) The protein expression of FOXO3a in shRNA-FOXO3a-infected C28/I2 cells was detected by western blotting assays. (E–K) Western blotting assays were used to detect the protein expression of Aggrecan, ADAMTS5, C/EBPβ, p-C/EBPβ, caspase3, cleaved-caspase3, and FOXO3a in LV-sh-FOXO3a-infected C28/I2 cells treated with IL-1β. (L-M) The apoptosis rate was determined by a flow cytometer.
5). Comparison of Curative Effect of Human Umbilical Cord-Derived Mesenchymal Stem Cells and Their Small Extracellular Vesicles in Treating Osteoarthritis. International journal of nanomedicine, 2021 (PubMed: 34938076) [IF=6.6]

Application: IHC    Species: Human    Sample:

Figure 4. Exploration of the effects of two treatments on cartilage matrix in vivo. (A and B) The immunohistochemical staining was performed to clarify the expression of Collagen II, MMP13 and ADAMTS5 in articulate (The magnification in A is 100×, scale bar = 100 μm; in B is 400×, scale bar = 20 μm;). (C) Statistical analysis of the percentage of pixels in the image or selection in three groups. Data are expressed as mean ±SEM. Different number of asterisk (*) show significant differences between groups. (*= P
6). Targeting Parkin-regulated metabolomic change in cartilage in the treatment of osteoarthritis. iScience, 2024 (PubMed: 39220257) [IF=5.8]

Application: WB    Species: Mouse    Sample:

Figure 6. LLC inhibited NF-κB pathway and alleviated cartilage deterioration See also Figure S5. (A) GSEA analysis revealed that chemokine related pathways were enriched in aged cartilage while de-enriched in Prkn-KO cartilage. (B) mRNA level of genes associated with cartilage matrix, inflammation and matrix degrading enzymes of chondrocytes treated with LLC (n = 3). (C and D) Protein expression and (D) quantitative results of matrix degrading enzymes of chondrocytes treated with LLC (n = 3). (E and F) A representative image of immunohistochemistry staining and (F) quantitative results of the LLC-treated OA mouse model 4 weeks postoperatively (n = 3). Scale bar: 100 μm. (G and H) Protein expression and (H) quantitative results of IκBα, p-IKKα/β and IKKα/β of chondrocytes treated with LLC (n = 3). (I and J) Cytoplasmic and nuclear protein expression and (J) quantitative results of p65 of chondrocytes treated with LLC (n = 3). Statistical analysis was performed by two-tailed Student’s t test for comparisons of two groups.
7). The Protective Effect of Evodiamine in Osteoarthritis: An In Vitro and In Vivo Study in Mice Model. Frontiers in Pharmacology, 2022 (PubMed: 35795554) [IF=5.6]

Application: WB    Species: Mouse    Sample:

FIGURE 4 Evodiamine had a protective effect on the ECM degradation of IL-1β–induced chondrocytes. (A,B,C,D) Relative protein expressions of collagen-II, ADAMTS-5, and MMP-13 were quantitated by Western blot. (E,F) Collagen-II (green) was detected by immunofluorescence. Data were expressed as mean ± SD. Significant differences between different groups are indicated as ##p < 0.01, vs. control group; ∗∗ p < 0.01, vs. IL-1β alone stimulation group, n = 3.
8). Circular RNA CircFOXO3 Functions as a Competitive Endogenous RNA for Acid-Sensing Ion Channel Subunit 1 Mediating Oxeiptosis in Nucleus Pulposus. Biomedicines, 2024 (PubMed: 38540291) [IF=4.7]
9). Medical Prospect of Melatonin in the Intervertebral Disc Degeneration through Inhibiting M1-Type Macrophage Polarization via SIRT1/Notch Signaling Pathway. Biomedicines, 2023 (PubMed: 37371708) [IF=4.7]

Application: WB    Species: Human    Sample: NP cells

Figure 4 Protection on NP cells against the CM-induced inflammation and imbalanced ECM homeostasis by the treatment of MLT. NP cells were treated with different CMs collected from the LPS-stimulated RAW 264.7 Mφs in the presence or absence of MLT for 24 h, then followed by different analysis. (A) The relative mRNA levels of pro-inflammation cytokines in NP cells were measured by RT-qPCR analyses (n = 3). The levels of IL-6, TNF-α, iNOS and COX-2 mRNA declined in the presence of MLT treatment to Mφs at different dosages. (B) The relative mRNA levels of matrix anabolic and degrading enzymes in NP cells were measured by RT-qPCR analyses (n = 3). The levels of COL2A1 and TIMP-1 mRNA were upregulated, while the levels of MMP-13, ADAMTS4 and ADAMTS5 mRNA were downregulated in the presence of MLT treatment to Mφs at different dosages. (C,D) The relative protein levels of pro-inflammation cytokines in NP cells were measured by Western blot analyses (n = 3). (C) Western blotting showed the decline of inflammation-associated proteins. (D) Quantitative analysis showed the reduced levels of IL-6, TNF-α, iNOS and COX-2 protein after treatment with MLT to Mφs at different dosages. (E,F) The relative protein levels of matrix anabolic and degrading enzymes in NP cells were measured by Western blot analyses (n = 3). (E) Western blotting showed the different changes of ECM protein productions in NP cells. (F) Quantitative analysis showed the increased levels of COL2A1 and TIMP-1 protein, and the reduced levels of MMP-13, ADAMTS4 and ADAMTS5 protein after the treatment with MLT to Mφs at different dosages. Data are expressed as mean ± standard deviation. The two groups among the five groups are compared by using an independent t-test.
10). Casticin ameliorates osteoarthritic cartilage damage in rats through PI3K/AKT/HIF-1α signaling. Chemico-biological interactions, 2024 (PubMed: 38309612) [IF=4.7]
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