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:

CD63-mediated cloaking of VEGF in small extracellular vesicles contributes to anti-VEGF therapy resistance
Ma, S., et al. Cell Rep. 2021 Sep 28;36(13):109684. doi: 10.1016/j.celrep.2021.109684. PMID: 34586478
Cells were stained with anti-CD63 (1:200, mouse IgG2a isotype control, Santa Cruz Biotechnology) for 1 h at 4°C. Mouse IgG2a isotype control antibody was used to assess nonspecific binding in flow cytometry. Tumor tissues were harvested and fixed in 4% paraformaldehyde for immunohistochemistry. The expression of CD63 was significantly upregulated in bevacizumab-resistant tumors compared to sensitive ones. Small extracellular vesicles isolated from resistant cells showed increased VEGF cloaking.
Tags: activity of Mouse IgG2a Isotype Control; Mouse IgG2a Isotype Control

Inhibition of interferon-signalling halts cancer-associated fibroblast-dependent protection of breast cancer cells from chemotherapy
Broad, R. V., et al. Br J Cancer. 2021 Feb;124(4):811-822. doi: 10.1038/s41416-020-01186-8. PMID: 33318582
Mouse IgG2a isotype control was administered intraperitoneally to control groups at the same dose and schedule as the anti-IFNAR antibody. Treatment with ruxolitinib significantly reduced CAF-mediated protection of breast cancer cells from paclitaxel. Cancer-associated fibroblasts (CAFs) were isolated from patient-derived breast tumor samples. Interferon signaling in CAFs promotes resistance to chemotherapy in co-culture models. Inhibition of JAK1/2 restored sensitivity to paclitaxel in CAF-tumor cell co-cultures.
Tags: Mouse IgG2a Isotype Control mAb; Mouse IgG2a Isotype Control antibody for animal model

The immunity of the recombinant prokaryotic and eukaryotic subunit vaccines against cutaneous leishmaniasis
Salari, S., et al. PLoS Negl Trop Dis. 2021 Jan 21;15(1):e0009029. doi: 10.1371/journal.pntd.0009029. PMID: 33461006
Mice were immunized with recombinant proteins adjuvanted with CpG and then challenged with L. major promastigotes. Mouse IgG2a isotype levels were measured by ELISA to assess Th1 response. The eukaryotic vaccine induced higher IgG2a titers compared to the prokaryotic one. Protection against cutaneous leishmaniasis was evaluated by lesion size measurement. Both vaccines reduced parasite burden in the footpad.
Tags: Mouse IgG2a Isotype Control antibody for mouse tumor model; Mouse IgG2a Isotype Control for animal model

The functional curcumin liposomes induce apoptosis in C6 glioblastoma cells and C6 glioblastoma stem cells in vitro and in animals
Wang, Y., et al. Int J Nanomedicine. 2017 Feb 28;12:811-827. doi: 10.2147/IJN.S124899. PMID: 28280391
Curcumin liposomes were injected into C6 glioma-bearing rats. Mouse IgG2a isotype control was used in immunohistochemistry for nonspecific staining. Tumor volumes were significantly reduced in the treatment group. Apoptosis in glioblastoma stem cells was confirmed by TUNEL assay. Biodistribution showed enhanced accumulation in brain tumors.
Tags: Mouse IgG2a Isotype Control for mouse tumor model; Mouse IgG2a Isotype Control dose

Regeneration of the pulmonary vascular endothelium after viral pneumonia requires COUP-TF2
Zhao, G., et al. Sci Adv. 2021 May 14;7(20):eabg4369. doi: 10.1126/sciadv.abg4369. PMID: 33979496
Lung endothelial cells were sorted using anti-CD31 antibody with mouse IgG2a isotype control for gating. COUP-TF2 knockout impaired endothelial regeneration post-influenza infection. Single-cell RNA sequencing revealed altered transcriptional profiles in endothelial clusters. Viral pneumonia was induced by intranasal H1N1 administration. Wild-type mice showed restored vascular integrity by day 14 post-infection.
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