Mouse IgG2a Isotype Control Antibody

Mouse IgG2a isotype control and mutants (natural and engineered mIgG2a isotype controls) are useful for in vitro and in vivo studies from ELISA, Western blot (WB), to cell-based assay to animal model study. Mouse IgG2a antibodies are similar to human IgG1 antibodies in many aspects, thus researchers like to use the isotype of mouse IgG2a in mouse studies. Most of the therapeutic antibody drugs carry the natural human IgG1 constant regions. More and more engineered mutants in the human IgG1 Fc fragment have been tested during therapeutic antibody drug development. The corresponding mutations have been introduced to mouse IgG2a to study enhancement and abrogation effects of Fc binding. We generated mIgG2a isotype control mutants, the same mutations as those in the therapeutic antibodies and candidates because such mouse IgG2a isotype control mutants are needed when clients test their engineered antibody candidates. For example, P329G substitution combined with L234A L235A (mIgG2a LALAPG, Fc silenced) was shown to decrease effector functions by disrupting the Fc/Fcγ receptor interface.

The ideal recombinant mouse IgG2a isotype control has low or no specific binding to any mouse sample, and completely mouse variable regions and the mouse IgG2a kappa constant regions. The mouse IgG2a kappa isotype control and mutants can be used in ELISA, Western blot (WB), the cell-based assay or animal model as the antibody negative control.

Please contact us to ask for a quote for the engineered mouse IgG2a isotype control mutants, mIgG2a isotype control and mutants with Avi-, His-, and DYKDDDDK-tags, and biotinylated mIgG2a isotype control and mutants. A variety of conjugates (such as dyes and fluorophores) with the mouse IgG2a isotype control and mutants are available.

PA007129: In Vivo Grade Recombinant Mouse IgG2a Isotype Control Antibody (mIgG2a Isotype Control)

The in vivo grade recombinant mouse IgG2a isotype control antibody (mIgG2a isotype control) was produced in mammalian cells.
Immunogen: N/A.
Clone: 12B9.
Isotype: mouse IgG2a, kappa.
Applications: an isotype-matched negative control for mouse IgG2a kappa antibodies used in ELISA, Western Blot (WB), Flow Cytometry (Flow), Immunoprecipitation (IP), Immunohistochemistry (Paraffin) (IHC (P)), Immunohistochemistry (Frozen) (IHC (F)), and in vivo animal model research.
Formulation: 0.2 μM filtered solution of 1x PBS.
Purity: >95% by SDS-PAGE under reducing conditions.
Endotoxin: Less than 1 EU/mg of protein as determined by LAL method.

PA007136: In Vivo Grade Recombinant Mouse IgG2a-D265A Isotype Control Antibody (mIgG2a D265A Isotype Control)

The in vivo grade recombinant mouse IgG2a-D265A isotype control mutant (mIgG2a D265A isotype control) was produced in mammalian cells.
Immunogen: N/A.
Clone: 12B9.
Isotype: mouse IgG2a, kappa.
Applications: an isotype-matched negative control for mouse IgG2a D265A kappa antibodies used in ELISA, Western Blot (WB), Flow Cytometry (Flow), Immunoprecipitation (IP), Immunohistochemistry (Paraffin) (IHC (P)), Immunohistochemistry (Frozen) (IHC (F)), and in vivo animal model research.
Formulation: 0.2 μM filtered solution of 1x PBS.
Purity: >95% by SDS-PAGE under reducing conditions.
Endotoxin: Less than 1 EU/mg of protein as determined by LAL method.

PA007137: In Vivo Grade Recombinant Mouse IgG2a-N297A Isotype Control Antibody (mIgG2a N297A Isotype Control)

The in vivo grade recombinant mouse IgG2a-N297A isotype control mutant (mIgG2a N297A isotype control) was produced in mammalian cells.
Immunogen: N/A.
Clone: 12B9.
Isotype: mouse IgG2a, kappa.
Applications: an isotype-matched negative control for mouse IgG2a N297A kappa antibodies used in ELISA, Western Blot (WB), Flow Cytometry (Flow), Immunoprecipitation (IP), Immunohistochemistry (Paraffin) (IHC (P)), Immunohistochemistry (Frozen) (IHC (F)), and in vivo animal model research.
Formulation: 0.2 μM filtered solution of 1x PBS.
Purity: >95% by SDS-PAGE under reducing conditions.
Endotoxin: Less than 1 EU/mg of protein as determined by LAL method.

PA007138: In Vivo Grade Recombinant Mouse IgG2a-L234A L235A P329G Isotype Control Antibody (mIgG2a LALAPG Isotype Control)

The in vivo grade recombinant mouse IgG2a-L234A L235A P329G (mIgG1 LALAPG, Fc silenced) isotype control mutant (mIgG2a LALAPG isotype control) was produced in mammalian cells.
Immunogen: N/A.
Clone: 12B9.
Isotype: mouse IgG2a, kappa.
Applications: an isotype-matched negative control for mouse IgG2a LALAPG kappa antibodies used in ELISA, Western Blot (WB), Flow Cytometry (Flow), Immunoprecipitation (IP), Immunohistochemistry (Paraffin) (IHC (P)), Immunohistochemistry (Frozen) (IHC (F)), and in vivo animal model research.
Formulation: 0.2 μM filtered solution of 1x PBS.
Purity: >95% by SDS-PAGE under reducing conditions.
Endotoxin: Less than 1 EU/mg of protein as determined by LAL method.

Shipping: The product is shipped with ice pack. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
1 month from date of receipt, 2 to 8°C as supplied.
3 months from date of receipt, -20°C to -70°C as supplied.

Background

Mouse IgG antibodies are classified to four subclasses mainly based on the isotype of the heavy chain (IgG1, IgG2a or IgG2c, IgG2b, and IgG3 for the heavy chain; kappa and lamda for the light chain), of which IgG1 and kappa are the most abundantnamed in serum. IgG antibodies are large molecules of about 150 kDa composed of four peptide chains, two identical heavy chains of about 50 kDa and two identical light chains of about 25 kDa. The four peptides are arranged in a Y-shape tetramer by disulfide bonds formed between the two heavy chains and between a heavy chain and a light chain.

References of Mouse IgG2a Isotype Control Antibody:

Targeting KDM4 family epigenetically triggers antitumour immunity via enhancing tumour-intrinsic innate sensing and immunogenicity.
Sun, M., et al. Clin Transl Med. 2024 Feb;14(2):e1598. doi: 10.1002/ctm2.1598. PMID: 38390756
Anti-CD8 antibody, anti-IFNAR-1 antibody, mouse IgG2a isotype control, rat IgG2b isotype control and anti-PD-L1 antibody were purchased from BioXcell and applied for in vivo assay. ...To discover novel drugs capable of inducing type I IFN secretion, we performed a high-throughput screen using a commercial anti-cancer compound library obtained from TargeMol. ...Despite the remarkable clinical efficacy of cancer immunotherapy, considerable patients fail to benefit from it due to primary or acquired resistance. ...Herein, we demonstrated that KDM4-targeting chemotherapeutic drug JIB-04, epigenetically triggered the tumour-intrinsic innate immune responses and immunogenic cell death (ICD), resulting in impressive antitumour effects. ...Furthermore, JIB-04 induced adaptive resistance through the upregulation of programmed death-ligand 1 (PD-L1), which could be overcome with additional PD-L1 blockade.
Tags: activity of Mouse IgG2a Isotype Control; Mouse IgG2a Isotype Control

Protective interplay: Mycobacterium tuberculosis diminishes SARS-CoV-2 severity through innate immune priming.
Williams, B. D., et al. Front Immunol. 2024 Jun 20;15:1424374. doi: 10.3389/fimmu.2024.1424374. PMID: 38966641
Primary infection of angiotensin converting enzyme-2 (ACE2)-expressing airway and alveolar epithelial cells by SARS-CoV-2 initiates viral replication, pyroptosis of host cells, and activation of innate immune pathways. ...Several pattern recognition receptors (PRRs) detect SARS-CoV-2 and initiate innate responses, including endosomal toll-like receptor 3 (TLR3) and toll-like receptor 7 (TLR7) signaling pathways, as well as cytoplasmic RNA sensor, melanoma differentiation-associated protein 5 (MDA5). ...Upon activation of the PRRs, downstream signaling results in the production of antiviral interferons (IFNs), and cytokines and chemokines which create an anti-viral environment and recruits innate cells to the site of infection. ...In contrast to the acute hyperinflammatory profile associated with SARS-CoV-2, chronic M.tb infection is known to elicit a diverse array of proinflammatory and regulatory responses. ...Further insights can be gleaned from other bacterial and viral co-infection studies.
Tags: Mouse IgG2a Isotype Control mAb; Mouse IgG2a Isotype Control antibody for animal model

RNA structures within Venezuelan equine encephalitis virus E1 alter macrophage replication fitness and contribute to viral emergence.
Hickson, S. E., et al. PLoS Pathog. 2024 Sep 27;20(9):e1012179. doi: 10.1371/journal.ppat.1012179. PMID: 39331659
Raw264.7 cells were seeded 24h prior to infection. One hour prior to infection, the cells were pretreated with 10μg of mouse IgG2a isotype control or IFNAR1 Monoclonal Antibody, infected with TC83 or TC83/E1ID-syn at an MOI of 0.1 in the presence of antibody. ...Infectious virus from cell culture supernatants harvested at 10 and 22 hpi was titered by FFA. ...Thereafter, lysates were centrifuged at 16,000xg for 20 mins at 4°C and supernatants were transferred to a new tube. 50μl of Dynabeads protein G were washed 2x in 500μl lysis buffer, after which they were incubated in 200μl lysis buffer along with 12μg of mouse J2 IgG2a Or mouse IgG2a isotype control for 30 mins at RT. ...This was repeated 3x to increase modifications. Unmodified samples were similarly treated with DMSO. ...VEEV translation reporters were constructed based on previously described constructs.
Tags: Mouse IgG2a Isotype Control antibody for mouse tumor model; Mouse IgG2a Isotype Control for animal model

Gestational diabetes augments group B Streptococcus infection by disrupting maternal immunity and the vaginal microbiota.
Mercado-Evans, V., et al. Nat Commun. 2024 Feb 3;15(1):1035. doi: 10.1038/s41467-024-45336-6. PMID: 38310089
For NK cell depletion experiments, mice received intraperitoneal injections of 250 μg of anti-NK1.1 antibody, or 250 μg of mouse IgG2a isotype control on E13.5 and E15.5 in 100 μL. ...Mice were challenged with vaginal inoculations of GBS on E14.5 and E15.5 and sacrificed on E17.5 to assess in utero bacterial dissemination as described above. ...Vaginal and uterine tissues (E17.5) from GBS-challenged GDM and pregnant control mice were dissected and placed in tubes containing 500 μL RNA Protect and 1.0 mm zirconia/silica beads. ...To characterize the murine vaginal microbiota throughout healthy and gestational diabetic pregnancy, we serially sampled the vaginal lumen every three days beginning seven days before mating until day of GBS challenge. ...These PBS blanks were used as controls for environmental and sequencing contamination.
Tags: Mouse IgG2a Isotype Control for mouse tumor model; Mouse IgG2a Isotype Control dose

A PD-1-targeted, receptor-masked IL-2 immunocytokine that engages IL-2Rα strengthens T cell-mediated anti-tumor therapies.
Wu, J., et al. Cell Rep Med. 2024 Oct 15;5(10):101747. doi: 10.1016/j.xcrm.2024.101747. PMID: 39326410
For immune cell depletion studies, mAbs used were as follows: mouse IgG2a isotype control (300μg) clone C1.18.4, rat IgG2b isotype control (500μg), clone LFT-2, anti-NK1.1 (300μg) clone PK136, anti-CD4 (500μg) clone GK1.5, anti-CD8 (300μg) clone 2.43. ...To study the combination of PD1-IL2Ra-IL2 and CAR T cell therapy, huCD3/huMUC16 knock-in mice were lymphodepleted via a sublethal dose of total body irradiation (400 cGy) one day before subcutaneous implantation with 1x107 ID8-VEGF/huMUC16Δ tumor cells in the right flank. ...For flow cytometry analysis of lymphocyte populations, blood, spleens and tumors from treated mice were collected on indicated days. ...Tumors were excised and weighed before mincing. ...To prepare TIL samples for single cell RNA-seq and TCRseq, human PD-1 knock-in mice were subcutaneously implanted with MC38 tumor cells on both left and right flanks.
Tags: Mouse IgG2a Isotype Control antibody mouse dose range; bulk Mouse IgG2a Isotype Control

For more references about Mouse IgG2a Isotype Control Antibody please contact our scientific support team with message@sydlabs.com.

Questions and Answers:

Question: The monoclonal antibody produced by the hybridoma C1.18.4 (clone number) is used as a mouse IgG2a kappa isotype control. The antibody variable regions of MOPC21 (hybridoma clone number, isotype: mouse IgG1, kappa) are used to make a recombinant mouse IgG2a kappa isotype control. Both C1.18.4 and MOPC21 antibodies have unknown binding specificity.  Is your isotype control the C1.18.4 or MOPC21 antibody?
Answer: Our recombinant mouse IgG1 isotype controls (catalog # PA007129 and the mutants) are not from C1.18.4 or MOPC21 though we also produce and sell the recombinant C1.18.4 and MOPC21 mouse IgG2a isotype controls. Please contact us and ask for C1.18.4 and MOPC21 derived recombinant mouse IgG2a kappa isotype controls if you are interested.

Question: What is the advantage of the recombinant antibody production method compared to the other antibody production method(s)?
Answer: We use chemically defined medium for cell culturing to produce the recombinant antibody. There is significant advantage of the in vitro method. First, we do not use mouse ascites for antibody production. Second, although antibodies can be produced by cell culturing from over 90% of hybridomas, there is a finite and significant failure rate. The recombinant method can be of help. Third, some companies and scientists add FBS into hybridoma cell culturing medium, which results in antibody contamination. There is no such contamination in production of recombinant antibodies. Fourth, we can easily switch the isotype with different antibody constant regions, and generate mutations such as D265A which is used in therapeutic antibody research. It is better to use the same isotype control mutation.

Question: What is an isotype control antibody and why is it used?
Answer: An ideal isotype control matches the isotype of the primary antibody used in the application but lacks binding to the target. It is normally used as a negative control to differentiate any non-specific background signal from the specific antibody signal. Mouse IgG1 kappa isotype controls against KLH, TNP, DNP,  GST, HEL, His-tag, DYKDDDDK, c-myc, HA, and GFP meet the requirements in some cases but not in specific cases in the presence of the specific targets or cross-reactivity.

Other in vivo grade Recombinant IgG Isotype Control Antibodies and Mutants:
In vivo Grade Recombinant Human IgG1 Isotype Control Antibody and Mutants
In vivo Grade Recombinant Human IgG2 Isotype Control Antibody
In vivo Grade Recombinant Human IgG3 Isotype Control Antibody
In vivo Grade Recombinant Human IgG4-S228P Isotype Control Antibody and Mutants
In vivo Grade Recombinant Mouse IgG1 Isotype Control Antibody and Mutants
In vivo Grade Recombinant Mouse IgG2a Isotype Control Antibody and Mutants
In vivo Grade Recombinant Mouse IgG2b Isotype Control Antibody and Mutants
In vivo Grade Recombinant Mouse IgG2c Isotype Control Antibody and Mutants
In vivo Grade Recombinant Mouse IgG3 Isotype Control Antibody
In vivo Grade Recombinant Rat IgG1 Isotype Control Antibody
In vivo Grade Recombinant Rat IgG2a Isotype Control Antibody
In vivo Grade Recombinant Rat IgG2b Isotype Control Antibody
In vivo Grade Recombinant Rat IgG2c Isotype Control Antibody
In vivo Grade Recombinant Hamster IgG1 Isotype Control Antibody
In vivo Grade Recombinant Hamster IgG2 Isotype Control Antibody

In vivo Grade Recombinant IgG Fc Proteins:
In vivo Grade Recombinant Human IgG1 Fc Protein (hIgG1)
In vivo Grade Recombinant Human IgG2 Fc Protein (hIgG2)
In vivo Grade Recombinant Human IgG4 Fc Protein (hIgG4)
In vivo Grade Recombinant Mouse IgG1 Fc Protein (mIgG1)
In vivo Grade Recombinant Mouse IgG2a Fc Protein (mIgG2a)
In vivo Grade Recombinant Mouse IgG2b Fc Protein (mIgG2b)
In vivo Grade Recombinant Rat IgG2a Fc Protein (rtIgG2a)
In vivo Grade Recombinant Rat IgG2b Fc Protein (rtIgG2b)
In vivo Grade Recombinant Llama IgG2b Fc Protein (lIgG2b)
In vivo Grade Recombinant Rabbit IgG Fc Protein (rIgG)

Fc ELISA Kits and Reagents:
Human Fc ELISA Kit
Mouse Fc ELISA Kit
Human Fc ELISA Reagent Kit
Mouse Fc ELISA Reagent Kit

Protein A, Resin, Columns, and ELISA Kits:
Recombinant Protein A
Protein A Affinity Resin
Pre-packed Protein A Column
Protein A ELISA Kit