Murine Anti-mIL-1β mAb

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 mAb against murine IL-1 beta

Neutralizing monoclonal antibody against murine IL-1β

InvivoGen provides a recombinant anti-mIL-1β-mIgG1 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].

 More details

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.