Recombinant anti-mouse IL-1β antibody

Notification: Our anti-mouse IL-1β antibody raised in mice and produced in hybridoma (mabg-mil1b/mabg-mil1b-5) has been removed from our catalog. Its coding sequence has been determined and the antibody is now produced by recombinant technology and purified from CHO cells.

Recombinant mouse IL-1 beta antibody

Neutralizing monoclonal antibody against murine IL-1β

InvivoGen provides a recombinant fully mouse anti-mouse IL-1β monoclonal antibody (mAb) that was previously extracted from hybridoma. It is now expressed and produced in Chinese hamster ovary (CHO) cells, ensuring reliability and lot-to-lot reproducibility. Thereby, common hybridoma-related drawbacks such as the generation of non-relevant mAbs containing aberrant light chains are avoided [1]. The sequence of the Anti-mIL-1β-mIgG1 is 100% murine (constant and variable regions), as the original clone (clone 7E3) was raised in mice using a proprietary method. This feature allows for reduced immunogenicity and risks of fatal hypersensitivity reactions upon repeated mAb injections into mice [2-4].

InvivoGen provides this antibody in two grades:

• In vitro use: Anti-mIL-1β-mIgG1
• In vivo use: Anti-mIL-1β-mIgG1 InvivoFit™

All InvivoFit™ products are handled in a clean room, filter-sterilized, and tested for bacterial contaminants. Additionally, this grade guarantees a low levels of endotoxins (<1 EU/ml). The buffer formulation is specifically adapted for in vivo studies.

Key features:

• Potent and specific neutralizing activity against mIL-1β (see figure)
• Sequence is 100% murine
• Murine IgG1 isotype (constant region)
• Free from non-relevant mAbs found in hybridoma-based productions
• Produced in animal-free facilities and defined media
• Low aggregation <5%
• InvivoFit™ grade is available
   

Anti-mIL-1β-mIgG1 is designed to efficiently neutralize the biological activity of mIL-1β. Interleukin 1β (IL-1β) is a soluble pro-inflammatory cytokine that plays a critical role in the host’s response to infection and injury [5].

References:

1. Bradbury, A. et al. 2018. When monoclonal antibodies are not monospecific: Hybridomas frequently express additional functional variable regions. mAbs, 10(4), 539–546
2. Mall C. et al., 2016. Repeated PD-1/PD-L1 monoclonal antibody administration induces fatal xenogenic hypersensitivity reactions in a murine model of breast cancer. Onco Immunol. 5(2):e1075114.
3. Murphy, J.T. et al., 2014. Anaphylaxis caused by repetitive doses of a GITR agonist monoclonal antibody in mice. Blood. 123(14):2172-2180.
4. Belmar N.A. et al., 2017. Murinization and H chain isotype matching of Anti-GITR antibody DTA-1 reduces immunogenicity-mediated anaphylaxis in C57BL/6 mice. J Immunol. 198:4502-4512.
5. Dinarello CA., 2011. Interleukin-1 in the pathogenesis and treatment of inflammatory diseases. Blood. 117:3720–3732.

Figures

Anti-mIL-1β-mIgG1 InvivoFit™ is a potent neutralization antibody. Increasing concentrations of Anti-mIL-1β-mIgG1 InvivoFit™ were incubated for 30 minutes with the recombinant cytokine mIL-1β (50 ng/ml) prior to the addition of HEK-Blue™ IL-1β cells. After overnight incubation, SEAP activity in the cell culture supernatant was assessed using QUANTI-Blue™ Solution. Data are shown in percentage (%) of neutralization.

Specifications

Target: Murine IL-1β (mIL-1β)

Specificity: No cross-reactivity with human IL-1β

Clone: 7E3

Source: CHO cells

Isotype: Murine IgG1, kappa

Control: Murine IgG1 Isotype Control

Formulation of Anti-mIL-1β-mIgG1: Lyophilized from 0.2 µm filtered solution in a sodium phosphate buffer with glycine, saccharose, and stabilizing agents

Formulation of Anti-mIL-1β-mIgG1 InvivoFit™: Lyophilized from 0.2 µm filtered solution in a sodium phosphate buffer with 5% saccharose

Tested applications Blocking & Neutralization

Quality control:

• These products have been validated using neutralization cellular assays.
• The complete sequence of these antibodies has been verified.
• The absence of bacterial contamination (e.g. lipoproteins and endotoxins) has been confirmed using HEK-Blue™ TLR2 and HEK‑Blue™ TLR4 cells.
• The endotoxin level in Anti-mIL-1β-mIgG1 InvivoFit™ is <1 EU/mg (determined by a LAL assay).

Contents

Please note: each mAb is sold separately.

Anti-mIL-1β-mIgG1

• mil1b-mab9-02​: 200 µg, lyophilized

Anti-mIL-1β-mIgG1 InvivoFit™

• mil1b-mab9-1​: 1 mg, lyophilized
• mil1b-mab9-10​: 10 mg, lyophilized
   

 The product is shipped at room temperature.

 Store lyophilized antibody at -20 °C.

 Lyophilized product is stable for at least 1 year.

 Avoid repeated freeze-thaw cycles.

InvivoFit™

InvivoFit™ is a high-quality standard specifically adapted for in vivo studies. InvivoFit™ products are filter-sterilized (0.2 µm) and filled under strict aseptic conditions in a clean room. The level of bacterial contaminants (endotoxins and lipoproteins) in each lot is verified using a LAL assay and a TLR2 and TLR4 reporter assay.

Details

Interleukin-1 beta (IL-1β) is a potent pro-inflammatory cytokine involved in a variety of cellular activities including cell migration, T lymphocyte activation and pro-inflammatory cytokine production [1]. Its inactive precursor, pro-IL-1β, is secreted by immune cells, notably the innate system as well as epithelia cells in response to stimuli such as inflammation, infection or cell damage. In order to get activated, pro-IL-1β is cleaved by caspase-1 upon inflammasome activation [1,2].

Following caspase-1-dependent processing, the mature IL-1β is rapidly secreted through pores of the plasma membrane. There, it interacts with the IL-1 receptor 1 (IL-1R1), which is shared with IL-1α. Upon receptor-ligand binding, a structural change of the receptor occurs allowing the co-receptor IL-1R3 to bind and form a trimeric complex. Subsequently, the intracellular adapter protein MyD88 (myeloid differentiation primary response 88) becomes phosphorylated triggering an intricate sequence of (auto-) phosphorylation, complex formation and ubiquitination events. Finally, activated NK-kB, c-Jun N-terminal kinase (JNK) and p38 mitogen-activated protein kinase (MAPK) pathways induce the expression of inflammatory cytokines and chemokines, such as IL-6 and IL-8 [3]. Due to its role in mediating acute and chronic inflammation, IL-1β has emerged as a therapeutic target for auto-inflammatory diseases [4].

References:

1. O’Neill L., 2008. The interleukin-1 receptor/Toll-like receptor superfamily: 10 years of progress. Immunol. Rev. 226, 10–18. 4. 2.
2. Dinarello C., 2018. Overview of the IL-1 family in innate inflammation and acquired immunity. Immunol Rev. 281(1): 8–27.
3. Weber A. et al., 2010. Interleukin-1 (IL-1) pathway. Sci Signal. 3(105).
4. Dinarello CA., 2011. Interleukin-1 in the pathogenesis and treatment of inflammatory diseases. Blood. 117:3720–3732.