Human IgG1 Isotype Control Antibody

Different from polyclonal "human IgG1 isotype control antibodies" purified from human myeloma serum, the recombinant human IgG1 isotype control (hIgG1 kappa isotype control antibody) and mutants are produced in mammalian cells (HEK293 and CHO cells), thus are monoclonal and there is no potential contamination of any other antibody isotype(s). The recombinant human IgG1 isotype control has low or no specific binding to any human sample, and humanized or completely human variable regions in addition to the human IgG1 kappa constant regions. The 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 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 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 human IgG1 isotype control mutants, hIgG1 isotype control and silent Fc mutants with Avi-, His-, and DYKDDDDK-tags, and biotinylated hIgG1 isotype control and Fc silent mutants. A variety of conjugates (such as a fluorophore) with the human IgG1 isotype control and Fc effector-silent mutants are available.

PA007125: In Vivo Grade Recombinant Human IgG1 kappa Isotype Control Antibody (hIgG1 Isotype Control)

The in vivo grade recombinant human IgG1 kappa isotype control antibody (hIgG1 isotype control) was produced in CHO cells.
Immunogen: N/A.
Clone: 4F17.
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. Endotoxin ≤0.05 EU/mg for PA007125.05, in vivo grade recombinant human IgG1 isotype control antibody, premium grade (endotoxin ≤0.05 EU/mg).

PA007134: In Vivo Grade Recombinant Human IgG1-L234A L235A P329G (hIgG1 LALAPG) kappa Isotype Control Antibody (hIgG1 LALAPG Isotype Control)

The in vivo grade recombinant human IgG1-L234A L235A P329G (hIgG1 LALAPG, Fc silenced) kappa isotype control mutant (hIgG1 LALAPG isotype control) was produced in CHO cells.
Immunogen: N/A.
Clone: 4F17.
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. Endotoxin ≤0.05 EU/mg for PA007134.05, in vivo grade recombinant human IgG1-L234A L235A P329G (hIgG1 LALAPG) isotype control antibody, premium grade (endotoxin ≤0.05 EU/mg).

PA007133: In Vivo Grade Recombinant Human IgG1-N297A kappa Isotype Control Antibody (hIgG1 N297A Isotype Control)

The in vivo grade recombinant human IgG1-N297A kappa isotype control mutant (hIgG1 N297A isotype control) was produced in mammalian cells.
Immunogen: N/A.
Clone: 4F17.
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.

PA007132: In Vivo Grade Recombinant Human IgG1-D265A kappa Isotype Control Antibody (hIgG1 D265A Isotype Control)

The in vivo grade recombinant human IgG1-D265A kappa isotype control mutant (hIgG1 D265A isotype control) was produced in mammalian cells.
Immunogen: N/A.
Clone: 4F17.
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.

PA007139: In Vivo Grade Recombinant Human IgG1-D265A N297A kappa Isotype Control Antibody (hIgG1 DANA Isotype Control)

The in vivo grade recombinant human IgG1-D265A N297A (hIgG1 DANA) kappa isotype control mutant (hIgG1 DANA isotype control) was produced in mammalian cells.
Immunogen: N/A.
Clone: 4F17.
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.

PA007140: In Vivo Grade Recombinant Human IgG1-S267E L328F (hIgG1 SELF) kappa Isotype Control Antibody (hIgG1 SELF Isotype Control)

The in vivo grade recombinant human IgG1-S267E L328F (hIgG1 SELF, Fc activated) kappa isotype control mutant (hIgG1 SELF isotype control) was produced in mammalian cells.
Immunogen: N/A.
Clone: 4F17.
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.
3 months from date of receipt, -20°C to -70°C as supplied.

Background of Human IgG1 Isotype Control

Isotype control antibodies (negative control antibodies or reference antibodies) are frequently used in various antibody-related in vivo and in vitro experiments such as ELISA, Western blot, immunofluorescence, immunohistochemistry, immunoprecipitation, and flow cytometry. Isotype control antibodies are used as negative controls of primary antibodies to check presence of non-specific binding to Fc receptors on the cell surface, other cellular proteins, carbohydrates, and lipids. Thus, the isotype control antibody normally matches the host species and isotype, including light chains (such as human IgG1 kappa antibody), concentration, and conjugation format of the experimental primary antibody.

References of Human IgG1 Isotype Control Antibody:

Clinical Scaleup of Humanized AnnA1 Antibody Yielded Unexpected High Reticuloendothelial (RES) Uptake in Mice
Xu, L. L., et al. Antibodies (Basel). 2025 Feb 6;14(1):14. doi: 10.3390/antib14010014. PMID: 39982229
Four antibodies were compared in this study, which included human IgG1 isotype control (Cat #0151K-01, Southern Biotech, Birmingham, AL, USA), mouse version (mAnnA1) (PRISM, La Jolla, CA, USA), and two humanized versions (hAnnA1 and hAnnA1-mut). Biodistribution studies were performed 24 h after injection of Tc-99m-HYNIC radiolabeled antibodies (purity > 98%). Our results showed that Tc-99m-HYNIC-hAnnA1 had low spleen and liver retention not statistically different from Tc-99m-HYNIC-IgG1 and Tc-99m-HYNIC-mAnnA1, with corresponding higher blood levels; however, Tc-99m-HYNIC-hAnnA1-mut had high levels in the spleen and liver with differences identified among the mouse strains, radiolabeling conditions, and injection routes. The AnnA1 antibody serum binding assay was performed using NuPAGE Bis-Tris gel electrophoresis, as per the manufacturer’s instruction. These results showed that the hAnnA1-mut antibody was shifted upwards in the gel because of the higher molecular weight of sera proteins bound to the hAnnA1-mut.
Tags: activity of human IgG1 isotype control antibody; human IgG1 isotype control

S-531011, a Novel Anti-Human CCR8 Antibody, Induces Potent Antitumor Responses through Depletion of Tumor-Infiltrating CCR8-Expressing Regulatory T Cells
Nagira, Y., et al. Mol Cancer Ther. 2023 Sep 5;22(9):1063-1072. doi: 10.1158/1535-7163.MCT-22-0570. PMID: 37420296
In vitro ADCC assay against a human CCR8-expressing cell line To assess cell death mediated by natural killer (NK) cells in the presence of S-531011, human CCR8-expressing Rat-1 cells were labeled with Calcein-AM (Dojindo) at a final concentration of 10 μg/mL and incubated with human FcγRIIIA-expressing KHYG-1 cells at a ratio of 1:30 in the presence of S-531011 or human IgG1 isotype control antibody (Clone QA16A12, BioLegend) at 37°C for 90 minutes. For measurement of maximum or spontaneous calcein release, Triton X-100 solution or assay buffer was added without human FcγRIIIA-expressing KHYG-1 cells, respectively. S-531011 was administered as a single intravenous injection to the CT26.WT tumor-bearing hCCR8 KI mice at doses 0.05, 0.5, or 5 mg/kg. The proportion of Tregs was decreased by S-531011 in a dose-dependent manner, but that of Tconv cells was not affected. S-531011 depleted tumor-infiltrating CCR8⁺ Tregs and showed potent antitumor activity in a tumor-bearing hCCR8 KI mouse model.
Tags: human IgG1 recombinant isotype control; human IgG1 isotype control antibody for cancer research

Therapeutic efficacy of humanized monoclonal antibodies targeting dengue virus nonstructural protein 1 in the mouse model
Tien, S. M., et al. PLoS Pathog. 2022 Apr 29;18(4):e1010469. doi: 10.1371/journal.ppat.1010469. PMID: 35486576
The binding ability of various humanized mAbs including h2E8-69, h2E8-70, h33D2, isotype control human IgG1 (hIgG1), mouse 33D2 (m33D2), and isotype control mouse IgG2a (mIgG2a) to Huh7 cells infected with four different DENV serotypes were analyzed by flow cytometry. The results showed that treatment with 100 μg/mouse h2E8-69, h2E8-70 and h33D2 all caused a reduction of DENV-induced prolonged bleeding time (Fig 2B) and skin hemorrhage (Fig 2C and 2D) compared to mice treated with isotype control hIgG1. To evaluate h33D2 in NS1-induced permeability change, HMEC-1 cells (3 × 10^5/well) were seeded onto transwells for 48 h. For CDC assay of the LALAPG variant, Huh-7 cells were used as target cells. The results showed that h33D2 at 10 μg/ml caused significant NK cell cytotoxic effect against all four serotypes of DENV-infected Huh-7 cells, whereas h33D2-LALAPG did not cause NK cell cytotoxic effect compared with isotype control hIgG1 (Fig 7).
Tags: human IgG1 isotype control antibody for mouse tumor model; human IgG1 isotype control for animal model

Preclinical Characterization of a Novel Monoclonal Antibody NEO-201 for the Treatment of Human Carcinomas
Fantini, M., et al. Front Immunol. 2018 Jan 4;8:1899. doi: 10.3389/fimmu.2017.01899. PMID: 29354121
Target cells were then treated with 10 μg/mL of human IgG1 isotype control antibody (Thermo Fisher Scientific, Waltham, MA, USA) or NEO-201 antibody unless otherwise indicated, and then NK cells were added at effector-to-target (E:T) ratios of 12.5:1 and 25:1. NEO-201 exhibited broad reactivity against a range of human carcinoma cell lines and tumor tissues, but was not observed to bind the majority of healthy tissues. In addition, NEO-201 exhibited both ADCC and CDC activity against human carcinoma cells in vitro and largely attenuated the growth of human pancreatic xenograft tumors in vivo both alone and in combination with human peripheral blood mononuclear cells (PBMCs) as the effector cell source for ADCC. Flow cytometry analysis was used to profile a panel of human carcinoma cell lines for NEO-201 binding. Biodistribution studies were conducted utilizing radiolabeled NEO-201 in female and male NU/NU nude mice with established CFPAC-1 xenograft tumors.
Tags: human IgG1 isotype control for flow cytometry; human IgG1 isotype control for mouse tumor model

In vitro and in vivo properties of a fully human IgG1 monoclonal antibody that combats multidrug resistant Pseudomonas aeruginosa
Adawi, A., et al. Int J Mol Med. 2012 Sep;30(3):455-64. doi: 10.3892/ijmm.2012.1040. PMID: 22735858
A human IgG1 isotype control mAb (32) was used in a number of in vitro and in vivo studies. To further scrutinize the specificity of LST-007 binding and assess its capability to bind the ‘naturally-occurring’ flagellin type b, ELISA studies were established whereby the PA target was in the form of immobilized whole PA bacteria tethered to ELISA plates via poly-L-lysine (PLL). In these binding studies (Fig. 3), PA strains 27853 (type a flagellin), Pa01 (type b flagellin) and 2 additional flagellin type b containing PA strains Ka02 and 25619 (as determined by PCR analysis, data not shown) were immobilized on plates. Concentration-dependent inhibition profiles on PA motility was observed with either the ex vivo (Fig. 4A) or exogenous source (Fig. 4B) of LST-007 with complete suppression of PA motility observed at 6.6 nM (∼1 μg/ml) mAb. The specificity of this effect was confirmed since an irrelevant human isotype control mAb administered at 20 mg/kg twice at +1 and +25 h, failed to curtail mortality which essentially superimposed on the mortality profile observed with the FB treatment group (Fig. 6).
Tags: in vivo study of human IgG1 isotype control antibody; purified human IgG1 isotype control

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

The following mutations have been introduced to alter the constant regions of an IgG1 antibody to change its immunologic properties by Fc interaction with host proteins:

hIgG1 heavy chain R214K
hIgG1 heavy chain R214K/N297A
hIgG1 heavy chain L234A/L235A
hIgG1 heavy chain L234F/L235E/P331S
hIgG1 heavy chain L235A/G237A
hIgG1 heavy chain D356E/L358M
hIgG1 heavy chain F126L
hIgG1 heavy chain D265A
hIgG1 heavy chain F243L/R292P/Y300L/V305I/P396L
hIgG1 heavy chain S239D/I332E
hIgG1 heavy chain S239D/I332E/A330L
hIgG1 heavy chain S298A/E333A/K334A
hIgG1 heavy chain G236A/S239D/I332E
hIgG1 heavy chain K326W/E333S
hIgG1 heavy chain S267E/H268F/S324T
hIgG1 heavy chain E345R/E430G/S440Y
hIgG1 heavy chain N297A
hIgG1 heavy chain N297Q
hIgG1 heavy chain N297G
hIgG1 heavy chain L235E
hIgG1 heavy chain L234A/L235A
hIgG1 heavy chain M252Y/S254T/T256E
hIgG1 heavy chain M428L/N434S
hIgG1 heavy chain S267E/L328F
hIgG1 heavy chain N325S/L328F

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