Chimeric Human IgG1 Isotype Control Antibody

A chimeric human IgG1 isotype control consists of murine variable regions and human IgG1 constant regions. The chimeric human IgG1 kappa isotype control and mutants such as Fc-silenced can be used in ELISA, Western blot (WB), the cell-based assay or animal model as the antibody negative control.

A number of chimeric hIgG1 isotype control mutants are available. Most of the therapeutic antibody drugs carry the natural human IgG1 constant regions. There are some engineered mutants in the IgG1 Fc fragment. Changes to the IgG Fc sequence including glycosylation sites (such as N297A or N297G) have been used to modulate Fc functions. Even a single mutation may have both enhancement and abrogation effects on Fc binding though multiple mutation combinations are commonly used. We generated chimeric hIgG1 isotype control mutants, the same mutations as those in the therapeutic antibodies and candidates because such human IgG1 isotype control mutants are needed when clients test their engineered antibody candidates. For example, P329G substitution combined with L234A L235A (hIgG1 LALAPG, Fc silenced) was shown to decrease effector functions by disrupting the Fc/Fcγ receptor interface. The silent Fc effctor functions human IgG1 isotype controls are useful for in vivo use.

The isotype controls are normally used to differentiate any non-specific background signal from the specific antibody signal, including the immunogenicity if used in animals. Most in vivo functional grade human IgG antibodies are used in mice for in vivo studies. Besides the antibody Complementarity-Determining Regions (CDRs), two human or humanized variable regions including the one in the human or humanized IgG isotype control do not necessarily induce the same immune response in mice because in some cases, even one amino acid difference can make difference. Mouse variable regions can be used to show the potential immunogenicity induced by the human variable regions in the human or humanized IgG antibodies to test. Thus, in some cases, it is better to use the mouse variable region in the human IgG isotype controls. We also produce and sell recombinant human IgG isotype controls from the popuplar mouse hybridoma clones, MOPC21 and C1.18.4, as well as recombinant human IgG1 kappa isotype controls against KLH, TNP, DNP,  GST, HEL, His-tag, DYKDDDDK, c-myc, HA, GFP, and unknown specificity.

Please contact us to ask for a quote for the Fc silenced chimeric human IgG1 isotype control mutants, chimeric hIgG1 isotype control and silent Fc mutants with Avi-, His-, and DYKDDDDK-tags, and biotinylated chimeric hIgG1 isotype control and Fc silent mutants. A variety of conjugates (such as a fluorophore) with the chimeric human IgG1 isotype control and Fc effector-silent mutants are available.

PA007187: In Vivo Grade Recombinant Chimeric Human IgG1 kappa Isotype Control Antibody (Chimeric hIgG1 Isotype Control)

The in vivo grade recombinant chimeric human IgG1 kappa isotype control antibody (chimeric hIgG1 isotype control) was produced in CHO cells.
Immunogen: N/A.
Clone: 4F17m.
Isotype: human IgG1, kappa.
Applications: an isotype-matched negative control for human IgG1 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 Level: Less than 1 EU/mg of protein as determined by LAL method.

PA007188: In Vivo Grade Recombinant Chimeric Human IgG1-D265A kappa Isotype Control Antibody (Chimeric hIgG1 D265A Isotype Control)

The in vivo grade recombinant chimeric human IgG1-D265A kappa isotype control mutant (chimeric hIgG1 D265A isotype control) was produced in mammalian cells.
Immunogen: N/A.
Clone: 4F17m.
Isotype: human IgG1, kappa.
Applications: an isotype-matched negative control for human IgG1 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 Level: Less than 1 EU/mg of protein as determined by LAL method.

PA007189: In Vivo Grade Recombinant Chimeric Human IgG1-N297A kappa Isotype Control Antibody (Chimeric hIgG1 N297A Isotype Control)

The in vivo grade recombinant chimeric human IgG1-N297A kappa isotype control mutant (chimeric hIgG1 N297A isotype control) was produced in mammalian cells.
Immunogen: N/A.
Clone: 4F17m.
Isotype: human IgG1, kappa.
Applications: an isotype-matched negative control for human IgG1 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 Level: Less than 1 EU/mg of protein as determined by LAL method.

PA007190: In Vivo Grade Recombinant Chimeric Human IgG1-L234A L235A P329G (hIgG1 LALAPG) kappa Isotype Control Antibody (Chimeric hIgG1 LALAPG Isotype Control)

The in vivo grade recombinant chimeric human IgG1-L234A L235A P329G (hIgG1 LALAPG, Fc silenced) kappa isotype control mutant (chimeric hIgG1 LALAPG isotype control) was produced in CHO cells.
Immunogen: N/A.
Clone: 4F17m.
Isotype: human IgG1, kappa.
Applications: an isotype-matched negative control for human IgG1 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 Level: Less than 1 EU/mg of protein as determined by LAL method.

PA007191: In Vivo Grade Recombinant Chimeric Human IgG1-D265A N297A kappa Isotype Control Antibody (Chimeric hIgG1 DANA Isotype Control)

The in vivo grade recombinant chimeric human IgG1-D265A N297A (hIgG1 DANA) kappa isotype control mutant (chimeric hIgG1 DANA isotype control) was produced in mammalian cells.
Immunogen: N/A.
Clone: 4F17m.
Isotype: human IgG1, kappa.
Applications: an isotype-matched negative control for human IgG1 DANA 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 Level: Less than 1 EU/mg of protein as determined by LAL method.

PA007192: In Vivo Grade Recombinant Chimeric Human IgG1-S267E L328F (hIgG1 SELF) kappa Isotype Control Antibody (Chimeric hIgG1 SELF Isotype Control)

The in vivo grade recombinant chimeric human IgG1-S267E L328F (hIgG1 SELF, Fc activated) kappa isotype control mutant (chimeric hIgG1 SELF isotype control) was produced in mammalian cells.
Immunogen: N/A.
Clone: 4F17m.
Isotype: human IgG1, kappa.
Applications: an isotype-matched negative control for human IgG1 SELF 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 Level: 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.
12 months from date of receipt, -20°C to -70°C as supplied.

References of Chimeric Human IgG1 Isotype Control Antibody:

Development of β-Lactoglobulin-Specific Chimeric Human IgEκ Monoclonal Antibodies for In Vitro Safety Assessment of Whey Hydrolysates
Knipping, K., et al. PLOS ONE. 2014 Aug 14;9(8):e106025. doi: 10.1371/journal.pone.0106025. PMID: 25153697
As an isotype control, a chimeric human IgG1 isotype control antibody was used (BioLegend, San Diego, CA). The aim of this study was to develop chimeric human IgE monoclonal antibodies (mAbs) against bovine β-lactoglobulin (BLG), a major allergen in cow’s milk, to investigate the importance of IgE in the immune response against whey hydrolysates. The IgE mAbs were developed by immortalizing B-cells from individuals allergic to cow’s milk using Epstein-Barr virus (EBV). The allergenicity of whey hydrolysates was assessed by measuring mediator release from humanized rat basophilic leukemia (RBL) cells sensitized with these IgE mAbs. The results demonstrate that the developed IgE mAbs are specific for BLG and can be used to assess the residual allergenicity of whey hydrolysates.
Tags: activity of Chimeric Human IgG1 Isotype Control; activity of Chimeric Human IgG1 Isotype Control mAb

An immunoglobulin E-reactive chimeric human immunoglobulin G1 anti-idiotype inhibits basophil degranulation through cross-linking of FcεRI with FcγRIIb
Wigginton, S. J., et al. Clinical & Experimental Allergy. 2008 Feb;38(2):313-319. doi: 10.1111/j.1365-2222.2007.02896.x. PMID: 18070165
Chimeric human IgG1 isotype control (Biolegend, San Diego, CA, USA) was used as a negative control in all experiments. This study aimed to investigate whether a chimeric human IgG1 anti-idiotype could inhibit IgE-mediated basophil degranulation by cross-linking FcεRI with FcγRIIb. Basophils were pre-incubated with the chimeric anti-idiotype or control antibodies before allergen challenge. Degranulation was assessed by measuring histamine release using a fluorometric assay. The results suggest that co-engagement of FcεRI and FcγRIIb by the chimeric anti-idiotype significantly reduces basophil activation.
Tags: Chimeric Human IgG1 Isotype Control mAb; human IgG1 recombinant isotype control

Anti-CD20 (rituximab) treatment for canine B-cell lymphoma: a potential model for human B-cell lymphomas
Suter, S. E., et al. Frontiers in Veterinary Science. 2022 Jan 25;9:1085674. doi: 10.3389/fvets.2022.1085674. PMID: 36686180
A chimeric human IgG1 isotype control antibody (BioLegend) was used to assess non-specific binding in flow cytometry experiments. This study evaluates the efficacy of rituximab, a chimeric anti-CD20 antibody, in a canine model of B-cell lymphoma. Peripheral blood mononuclear cells (PBMCs) were isolated from dogs with confirmed B-cell lymphoma. Flow cytometry was used to confirm CD20 expression on lymphoma cells prior to treatment. Rituximab treatment resulted in a significant reduction in tumor burden in treated dogs compared to controls.
Tags: Chimeric Human IgG1 Isotype Control antibody for cancer research; Chimeric Human IgG1 Isotype Control antibody for flow cytometry

Targeting of the CD33-calicheamicin immunoconjugate Mylotarg (CMA-676) in acute myeloid leukemia: in vivo and in vitro saturation and internalization by leukemic and normal myeloid cells
van Der Velden, V. H., et al. Blood. 2001 May 15;97(10):3197-3204. doi: 10.1182/blood.V97.10.3197. PMID: 11342445
A chimeric human IgG1 isotype control was used to determine non-specific binding of the Mylotarg conjugate to AML cells. The study aimed to investigate the binding, internalization, and saturation kinetics of Mylotarg in acute myeloid leukemia (AML) cells. Bone marrow and peripheral blood samples were obtained from AML patients and healthy donors. Flow cytometry and confocal microscopy were used to assess CD33 expression and antibody internalization. Mylotarg showed specific binding to CD33-positive cells, with minimal binding to CD33-negative cells.
Tags: Chimeric Human IgG1 Isotype Control for animal model; Chimeric Human IgG1 Isotype Control for mouse tumor model

The N-terminal end of the CH2 domain of chimeric human IgG1 anti-HLA-DR is necessary for C1q, FcγRI and FcγRIII binding
Morgan, A., et al. Immunology. 1995 Oct;86(2):319-324. doi: Not available (pre-DOI era). PMID: 7490130
A chimeric human IgG1 isotype control was included to assess non-specific interactions with complement and Fc receptors. This study investigates the structural requirements for the binding of a chimeric human IgG1 anti-HLA-DR antibody to C1q, FcγRI, and FcγRIII. Mutants of the IgG1 antibody with deletions in the CH2 domain were generated to map binding sites. Binding assays were performed using purified human C1q and Fcγ receptors expressed on U937 cells. The results indicate that the N-terminal end of the CH2 domain is critical for effective binding to both complement and Fc receptors.
Tags: function of Chimeric Human IgG1 Isotype Control antibody; in vivo study of Chimeric Human IgG1 Isotype Control antibody

For more references about Chimeric Human IgG1 Isotype Control Antibody, please contact our scientific support team with message@sydlabs.com.

Syd Labs provides the following IgG Protein :
Human IgG1 Fc Protein
Human IgG1 Fc Protein
Human IgG2 Fc Protein
Human IgG3 Fc Protein
Human IgG4 Fc Protein
Llama IgG2b Fc Protein
Mouse IgG1 Fc Protein
Mouse IgG2a Fc Protein
Mouse IgG2b Fc Protein
Mouse IgG2c Fc Protein
Mouse IgG2C Protein
Mouse IgG3 Fc Protein
Rabbit IgG Protein
Rat IgG2a Fc Protein
Rat IgG2b Fc Protein

Syd Labs provides the following IgG Antibody :
Chimeric Human IgG1 Isotype Control Antibody
Chimeric Human IgG2 Isotype Control Antibody
Chimeric Human IgG4-S228P Isotype Control Antibody
Hamster IgG1 Isotype Control Antibody
Hamster IgG2 Isotype Control Antibody
Human IgG1 Isotype Control Antibody
Human IgG2 Isotype Control Antibody
Human IgG3 Isotype Control Antibody
Human IgG4-S228P Isotype Control Antibody
IgG Antibody
Mouse IgG1 Isotype Control Antibody
Mouse IgG2a Isotype Control Antibody
Mouse IgG2b Isotype Control Antibody
Mouse IgG2c Isotype Control Antibody
Mouse IgG3 Isotype Control Antibody
Rat IgG1 Isotype Control Antibody
Rat IgG2a Isotype Control Antibody
Rat IgG2b Isotype Control Antibody