Murine IgG1e3 Isotype Controls

Recombinant mouse IgG1e3 isotype control mAb against E. coli β-galactosidase

InvivoGen’s engineered Anti-β-Gal-mIgG1e3 antibody

InvivoGen provides a recombinant anti-β-Gal-mIgG1e3 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]. This control antibody targets E. coli's β-galactosidase (β-Gal), an irrelevant antigen that is not present in mice.

The sequence of Anti-β-Gal-mIgG1e3 is 100% murine (constant and variable regions), as the original clone (clone T9C6) was raised in mice using a proprietary method. Within the engineered murine IgG1e3 isotype, a point mutation D265A was introduced leading to a complete loss of unwanted Fc-associated effector functions [2]. Altogether, these alterations increase antibody performance and overcomes immunogenic events, especially upon repeated mAb injections into mice [3]. 

InvivoGen provides this antibody in two grades:

• In vitro use: Anti-β-Gal-mIgG1e3
• In vivo use: Anti-β-Gal-mIgG1e3 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:

• Targets E. coli β-galactosidase  antigen
• Sequence is 100% murine
• Murine IgG1e3 isotype (constant region)
• mIgG1e3 (IgG1 with a replacement of aspartic (D) acid by alanine (A) at position 265 point mutation) is effectorless
• 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

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. Baudino L. et al., 2008. Crucial role of aspartic acid at position 265 in the CH2 domain for murine IgG2a and IgG2b Fc-associated effector functions. J Immunol. 181(9):6664-9.
3. Yamazaki T. et al., 2005. Blockade of B7-H1 on macrophages suppresses CD4+ T cell proliferation by augmenting IFN-gamma-induced nitric oxide production. J Immunol. 175(3):1586-92.

Specifications

Target: E. coli β-galactosidase (β-Gal)

Specificity: No cross-reaction with murine proteins

Clone: T9C6

Source: CHO cells

Isotype: Murine IgG1e3 (D265A mutation; no effector function), kappa

Formulation of Anti-β-Gal-mIgG1e3: Lyophilized from 0.2 µm filtered solution in a sodium phosphate buffer with glycine, saccharose, and stabilizing agents

Formulation of Anti-β-Gal-mIgG1e3 InvivoFit™: Lyophilized from 0.2 µm filtered solution in a sodium phosphate buffer with 5% saccharose

Application: Negative control for murine IgG1e3 antibodies

Quality control:

• The absence of binding was confirmed by flow cytometry.
• The complete sequence of this antibody 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-β-Gal-mIgG1e3 InvivoFit™ is <1 EU/mg (determined by a LAL assay).

Contents

Please note: each mAb is sold separately.

Anti-β-Gal-mIgG1e3

• bgal-mab15-02​: 200 µg, lyophilized

Anti-β-Gal-mIgG1e3 InvivoFit™

• bgal-mab15-1​: 1 mg, lyophilized
• bgal-mab15-10​: 10 mg, lyophilized
• bgal-mab15-50​: 50 mg (5 x 10mg), 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.