Anti-MMP8 [Nb14]
Catalogue Number: AB04110-34.11-ABA
| Manufacturer: | Vector Laboratories, Inc (ABA) |
| Type: | Recombinant Monoclonal |
| Alias: | MMP-8; Neutrophil collagenase; Matrix metalloproteinase-8; Matrix Metallo-Proteinase 8; Collagenase 2; PMNL-CL; PMNL collagenase; MNL-CL; CLG1; HNC; Nb 70; Nanobody 70 |
| Shipping Condition: | Blue Ice |
| Unit(s): | 100 ug |
| Host name: | Camelid |
| Clone: | Nb14 |
| Isotype: | |
| Immunogen: | The original antibody was generated by immunizing an alpaca (Vicugna pacos) with the recombinant mouse catalytic domain of matrix metalloproteinase 8 (mMMP8_CD). A VHH library was constructed using total RNA from peripheral blood lymphocytes, and phages displaying the VHH repertoire were screened for mMMP8_CD-specific nanobodies. |
| Application: | ELISA, FA, InVivoA, SPR |
Additional Text
Gene Name
MMP8
Uniprot ID
O70138
Purification
Purified
Gene ID
17394
Antibody Clonality
Recombinant Monoclonal
Storage Note
Store at 4⁰C for up to 3 months. For longer storage, aliquot and store at -20⁰C.
Application Notes
The binding affinity of this antibody's original format (VHH) to different substrates, including the catalytic domain of mouse MMP8 (mMMP8_CD), full-length mouse MMP8 (mMMP8_FL), and the catalytic domain human MMP8 (hMMP8_FL), was measured by ELISA. It had a KD of 0.24 nM with mMMP8_CD, a KD of 1.69 nM with mMMP8_FL, and a KD of 158.4 nM with hMMP8_CD. SPR analysis was conducted to measure its binding kinetics to mMMP8_CD; a KD of 1.334 +/- 0.137 nM was measured. Its in vivo pharmacokinetic properties were studied by injecting a single dose intraperitoneally into mice; blood samples were collected at different time points, and the nanobody's level in the mouse serum was detected using ELISA to evaluate its persistence and clearance in the bloodstream over time. This antibody's inhibitory capacity against MMP8 was investigated; it demonstrated the strongest inhibition of mMMP8_CD with an IC50 of 4.359 µM using DQ gelatin as a substrate and an IC50 value of 19.5 µM using DQ collagen type I. This antibody was used to create a bivalent nanobody by linking it to itself and to clone Nb Albumin using flexible [G4-S]3 linkers. The resulting nanobodies (Nb 14-Alb and Nb 14-Alb-14) were used in in vivo experiments for various applications (Demeestere et al., 2016; PMID: 26775809). The original format of this antibody was used to create another trivalent nanobody by linking it to clones Nb 70 and Nb Albumin using the same linkers. The resulting nanobody (Nb 70-Alb-14) was used in vivo to study the effects of inhibiting matrix MMP8 and TNFR1 on sepsis, using endotoxemia and CLP (cecal ligation and puncture; a surgical procedure commonly used in animal models to induce polymicrobial sepsis) experimental models (Steeland et al., 2018; PMID: 29095202).
AB04110-34.11 Datasheet
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