Mouse Anti-ADAMTS5 Recombinant Antibody (clone 7B4) (CAT#: PABL-005)

Recombinant Mouse Antibody (7B4) is capable of binding to ADAMTS-5.


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FuncS

Figure 1 ADAMTS-4 and ADAMTS-5 mAbs specifically bind to and potently inhibit aggrecanase activity.

Figure 1 ADAMTS-4 and ADAMTS-5 mAbs specifically bind to and potently inhibit aggrecanase activity.

(a) Selective binding of mAbs for ADAMTS-4 (7E8.1E3) and ADAMTS-5 (7B4.1E11) is demonstrated by immunocytochemistry using ADAMTS-4, ADAMTS-5 and Null BacMam transduced CHO cells. ADAMTS-5 specific domain mapping and selective binding to purified recombinant proteins is profiled by ELISA (b; 12F4.1H7) and DELFIA (c; 7E8.1E3 and d; 12F4.1H7) (Results shown are n = 2 independent experiments with two dependent replicates for each concentration ± 95% CI). Functional potency of ADAMTS-4 (e) and ADAMTS-5 (f) mAbs and non-selective small molecule (GSK579149) is quantified vs isotype controls (not shown) by measuring percent inhibition of selective aggrecanase-mediated ARGS neoepitope formation from purified bovine aggrecan substrate. IC50 values for each mAb and GSK571949 are shown (inset, e and f) in relation to isotype control mAbs and DMSO, respectively, which did not demonstrate any detectible inhibitory activity. Consistent with binding specificity, no inhibition of aggrecanase activity was observed in this assay for ADAMTS-4 with ADAMTS-5 mAbs and ADAMTS-5 with ADAMTS-4 mAbs (not shown).

Larkin, J., Lohr, T. A., Elefante, L., Shearin, J., Matico, R., Su, J. L., ... & Tran, P. B. (2015). Translational development of an ADAMTS-5 antibody for osteoarthritis disease modification. Osteoarthritis and cartilage, 23(8), 1254-1266.

Inhib

Figure 2 Potent and sustained inhibition of aggrecanolysis from human OA cartilage explants by ADAMTS-5 mAb treatment.

Figure 2 Potent and sustained inhibition of aggrecanolysis from human OA cartilage explants by ADAMTS-5 mAb treatment.

ADAMTS-5 mAb treatment inhibits ARGS neoepitope release from sequentially unstimulated and IL-1b/OSM stimulated human OA cartilage explants (a), whereas only modest inhibition is observed with ADAMTS-4 mAb treatment, preferentially under cytokine-stimulated conditions. Mean percent inhibition of mAbs is shown relative to isotype control mAb (0% inhibition, dashed lines in a and b) and GSK571949 (100% inhibition) treatment. Experiments were conducted with seven replicates for each treatment condition from eight timepoints across multiple 28-day experiments consisting of a 21 day unstimulated phase followed by a 7-day cytokine-stimulated phase. A total of 13–28 independent OA patient experiments were conducted for each treatment condition. ADAMTS-4 (7E8.1E3 or 7C71.H1) and ADAMTS-5 (12F4.1H7 or 7B4.1E11) mAb treatments are shown as compiled data associated with each mAb specificity. All mAb treatments were held constant at 670 nM throughout the experiment and GSK571949 was similarly held at 2 μM. (b) Dose-dependent inhibition of ARGS neoepitope release is evident with GSK2394002 treatment in the absence of exogenous cytokine stimulation using cartilage from individual OA donors with elevated pre-treatment ARGS neoepitope levels (n = 2–8 independent donor experiments per treatment concentration and each point represents mean % inhibition from 21 day unstimulated phase, as described in (a)). Plots (a and b) are presented in standard box-and-whisker format with Tukey outliers noted (>1.5 times IQR, filled symbols).(c) Linear regression analysis suggests the response to GSK2394002 treatment correlates with pre-treatment ARGS neoepitope levels. Sustained suppression of ARGS neoepitope release is observed with pulse-chase GSK2394002 treatment in relation to small molecule (GSK571949) in individual donor experiments using cultures with elevated pre-treatment ARGS neoepitope levels (n = 4; each timepoint and ±95% CI error bar represents mean of four independent donor experiments where seven biological replicates for each donor/timepoint was used to calculate a within donor mean for each timepoint) (d).

Larkin, J., Lohr, T. A., Elefante, L., Shearin, J., Matico, R., Su, J. L., ... & Tran, P. B. (2015). Translational development of an ADAMTS-5 antibody for osteoarthritis disease modification. Osteoarthritis and cartilage, 23(8), 1254-1266.

FuncS

Figure 3 ADAMTS-5 mAb treatment suppresses joint disease severity in a mouse model of OA.

Figure 3 ADAMTS-5 mAb treatment suppresses joint disease severity in a mouse model of OA.

Mice treated prophylactically with anti-ADAMTS-5 (1x/week, i.p. 10–16 mg/kg) were protected from developing (a) histologic cartilage degeneration (two independent studies shown, n = 8–10 animals/group each with two DMM knees/animal, although not all knees in study two could be scored due to isolated tissue processing errors and in one group loss of a single mouse between surgery and treatment commencement, as demonstrated by individual knee symbols in each group ranging from 15 to 19/group). Representative toluidine blue stained knee joint sections are shown for each study group. (b) Mice treated prophylactically with anti-ADAMTS5 mAb (1x/week, i.p. 10 mg/kg) were protected from developing mechanical allodynia through 8 weeks post DMM, ***P < 0.001 vs time 0, n = 12/group. Mean ± 95% CI. (c) Prophylactic treatment with anti-ADAMTS5 mAb also protected mice from macrophage infiltration (F4/80 marker) into the DRG at 8 weeks post DMM. Arrows indicate examples of infiltrating macrophages in the DMM control and Isotype control mAb (IgG1) conditions.

Larkin, J., Lohr, T. A., Elefante, L., Shearin, J., Matico, R., Su, J. L., ... & Tran, P. B. (2015). Translational development of an ADAMTS-5 antibody for osteoarthritis disease modification. Osteoarthritis and cartilage, 23(8), 1254-1266.


Specifications

  • Immunogen
  • Human ADAM metallopeptidase with thrombospondin type 1 motif, 5
  • Host Species
  • Mouse
  • Derivation
  • Mouse
  • Type
  • Mouse IgG1
  • Specificity
  • Human ADAMTS5
  • Species Reactivity
  • Human
  • Clone
  • 7B4
  • Applications
  • Inhib, WB, ELISA, FuncS

Product Property

  • Purity
  • >95% as determined by analysis by SDS-PAGE
  • Concentration
  • Please refer to the vial label for the specific concentration.
  • Buffer
  • PBS
  • Preservative
  • No preservatives
  • Storage
  • Centrifuge briefly prior to opening vial. Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C long term. Avoid repeated freeze/thaw cycles.

Applications

  • Application Notes
  • The antibody 7B4 has been reported in applications of Inhibition, Western Blot, Enzyme-linked Immunosorbent Assay, Functional Assay. It's recommended that the optimal antibody concentration, dilution, incubition time etc. be carefully titrated in specific assays.

Target

  • Alternative Names
  • ADAMTS5; ADAM metallopeptidase with thrombospondin type 1 motif; 5; ADMP-2; ADAM-TS5; ADAMTS-5; ADAMTS11; ADAM-TS 5; ADAMTS-11; ADAM-TS 11; A disintegrin and metalloproteinase with thrombospondin motifs 5; aggrecanase-2; a disintegrin and metalloproteinase with thrombospondin motifs 11; a disintegrin-like and metalloprotease (reprolysin type) with thrombospondin type 1 motif; 5 (aggrecanase-2)

Product Notes

This is a product of Creative Biolabs' Hi-Affi™ recombinant antibody portfolio, which has several benefits including:

• Increased sensitivity
• Confirmed specificity
• High repeatability
• Excellent batch-to-batch consistency
• Sustainable supply
• Animal-free production

See more details about Hi-Affi™ recombinant antibody benefits.

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Chimeric Antibody

CAT Product Name Application Type
TAB-0692CLV Anti-Human ADAMTS5 Recombinant Antibody (7B4) ICC, ELISA Chimeric antibody (mouse/human)

Humanized Antibody

CAT Product Name Application Type
TAB-0693CLV Humanized Anti-Human ADAMTS5 Recombinant Antibody (12F4.1H7) FuncS Humanized antibody

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