Catalog No.	PA007125
Product Name	Human IgG1 Isotype Control Antibody (Clone: 4F17) | PA007125
Data Sheet	Technical Data Sheet | Satety Data Sheet
Supplier Name	Syd Labs, Inc.
Brand Name	Syd Labs
Synonyms	hIgG1 Isotype Control, hIgG1 D265A N297A, IgG1 LALAPG, IgG1 SELF Mutants
Summary	The In Vivo Grade Recombinant Human IgG1 Isotype Control Antibody was produced in CHO cells.
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.
Immunogen	N/A.
Form Of Antibody	0.2 μM filtered solution of 1x PBS.
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).
Purity	>95% by SDS-PAGE under reducing conditions.
Shipping	Recombinant Human IgG1 Isotype Control Antibody(hIgG1 Isotype Control), In Vivo Grade 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.
Order Offline	Phone: 1-617-401-8149 Fax: 1-617-606-5022 Email: message@sydlabs.com Or leave a message with a formal purchase order (PO) Or credit card.

Description

PA007125: Recombinant Human IgG1 Isotype Control Antibody (Clone: 4F17) , In Vivo Grade

The in vivo grade recombinant human IgG1 kappa isotype control antibody (hIgG1 isotype control) was produced in CHO cells.

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, Flag, 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 Flag-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.

Background

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:

1、A mouse model for the preclinical evaluation of immunosuppressive effector functions of human isotypes. The human IgG1 isotype is superior to IgG3
Mocikat, R., et al. Transplantation. 1994 Feb;57(3):405-11. doi: 10.1097/00007890-199402150-00016. PMID: 8108876
“Chimeric mouse anti-Thy-1.2 antibody with the human IgG1 Fc part was found to be equally effective in preventing graft-versus-host disease mortality as the highly protective anti-Thy-1.2 mouse IgG2a isotype, while human IgG3 was far less effective. …Antibodies against T cells are widely used as immunosuppressive agents in clinical therapy. …As effector functions of chimeric or humanized anti-T cell antibodies cannot be predicted in vitro, we compared T cell-depleting effects of human isotypes in vivo with their immunosuppressive consequences in a mouse BMT model. …This system is based on chimeric antibodies with a mouse pan T cell specificity and human constant regions. …This was not predictable by measuring the degree of T cell depletion in peripheral blood. T cell depletion in lymph nodes, however, exactly reflected the results obtained in the BMT system.”

2、Selection of Vaccinia Virus-Neutralizing Antibody from a Phage-Display Human-Antibody Library
Shin, et al. J Microbiol Biotechnol. 2019 Apr 28;29(4):651-657. doi: 10.4014/jmb.1812.12024. PMID: 30856707
“The VH and VL of SC34 and SC212 obtained from library screening were converted to human IgG1 (mAb VC34 and mAb VC212) via insertion into the mammalian expression vectors pRC13 (VH) and pKC12 (VL), respectively. …Because the original form of pKC12 contained the constant region of the kappa chain, the lambda constant region was cloned from the cDNA of B lymphocytes. …Chimpanzee Fabs against the VACV B5 envelope protein were isolated from a phage display Fab library derived from the bone marrow of chimpanzees vaccinated with VACV and then converted to mAbs with human IgG1 heavy-chain constant regions (8AH8AL and 8AH7AL with Kd values of 0.2 and 0.7 nM respectively). …Chimpanzee Fabs against the VACV A33 envelope protein were isolated from the same chimpanzee Fab phage library described above, and then converted to mAbs with human IgG1 heavy-chain constant regions (6C,nd 12F with Kd values of 0.14–20 nM). …This study solved the disadvantages of human plasma-derived VIG production by isolating human mAbs capable of neutralizing VACV from a phage-display scFv library prepared from B cells of VACV-vaccinated volunteers.”

3、Bi-isotype immunoglobulins enhance antibody-mediated neutrophil activity against Plasmodium falciparum parasites
Ogwang, et al. Front Immunol. 2024 Apr 8;15:1360220. doi: 10.3389/fimmu.2024.1360220. PMID: 38650925
“These were subsequently cloned upstream of the human IgG1 expression vectors and co-transfected into Expi293F cells. A total of 35 human mAbs were successfully expressed as shown by dot blots detecting human IgG following expression. …We applied a published analytical approach to investigate the specificity of J31. This involved creating an A score that represented the number of standard deviations above the background mean fluorescence intensity (MFI) of all antigens on the chip. …Next, whether an Fc region cross isotype (IgG–IgA) consisting of IgG and IgA sequences would increase the ADRB activity of J31 was tested. …The J31 IgG–IgA variant showed a twofold enhanced ADRB activity compared to IgG1. …Two mAbs, CIS43LS and L9LS, that target the circumsporozoite protein have shown greater than 80% protection in phase 1 controlled human infection studies.”

4、Glycosylation is required for the neutralizing activity of human IgG1 antibodies against human rabies induced by pre-exposure prophylaxis
Koike, G., et al. Immunobiology. 2021 Mar;226(2):152058. doi: 10.1016/j.imbio.2021.152058. PMID: 33609912
“As changes in the antibody‘s carbohydrate chain can interfere with its effector functions, we compared the glycosylation patterns of both neutralizing and non-neutralizing IgG1 induced by pre-exposure prophylaxis to human rabies and analyzed their influence on in vitro antibody neutralizing activities. …The purified IgG1 showed an electrophoretic pattern compatible with human IgG. …Our results suggest that antibody glycosylation is important for neutralizing RABV in vitro, since neutralizing IgG1 has a different glycosylation profile than non-neutralizing IgG1. …According to the World Health Organization, rabies vaccines for human application are derived from inactivated and purified RABV grown either in tissue culture or in chicken or embryonated duck eggs. …Four subclasses (IgG1–4) of IgGs are found in humans, with IgG1 and IgG3 having the highest affinities for activating type I FcγRs.”

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

Other in vivo grade Recombinant IgG Isotype Control Antibodies and Mutants:
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

In vivo Grade Recombinant IgG Fc Proteins:
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), In vivo Grade

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

Human IgG1 Isotype Control (4F17) from: In Vivo Grade Recombinant Human IgG1 Isotype Control Antibody: PA007125 Syd Labs

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