Human IgG4-S228P Isotype Control Antibody

The ideal recombinant human IgG4 isotype control antibody (hIgG4 S228P kappa isotype control) has low or no specific binding to any human sample, and humanized or completely human variable regions in addition to the human IgG4 S228P kappa constant regions. The human IgG4 kappa isotype control and mutants can be used in the cell-based assay or animal model as the antibody negative control when evaluating fully human, humanized, or chimeric antibodies.

The hIgG4 isotype control mutants were generated because the corresponding mutations had been introduced to some human IgG4 therapeutic antibody candidates. For example, S228P, an engineered mutation in the core hinge region, is used to prevent Fab-arm exchange in vivo of human IgG4 molecules; P329G substitution combined with S228P and L235E was shown to decrease effector functions by disrupting the Fc/Fcγ receptor interface. When clients test their engineered antibody candidates, the silent Fc effctor function human IgG4 isotype controls are useful for in vivo use.

Welcome to contact us and ask for a quote for the engineered human IgG4 isotype control mutants, hIgG4 isotype control and Fc silenced mutants with Avi-, His-, and DYKDDDDK-tags, and biotinylated hIgG4 isotype control and silent Fc mutants. A variety of conjugates (such as dyes and fluorophores) with the human IgG4 isotype control and silent Fc effctor function mutants are available.

PA007128: In Vivo Grade Recombinant Human IgG4-S228P Isotype Control Antibody (hIgG4 Isotype Control)

The recombinant human IgG4-S228P kappa isotype control antibody (hIgG4 S228P isotype control) was produced in CHO cells.
Immunogen: Trinitrophenol (TNP).
Clone: 1R12.
Isotype: human IgG4, kappa.
Applications: an isotype-matched negative control for human IgG4 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 PA007128.05, in vivo grade recombinant human IgG4-S228P isotype control antibody, premium grade (endotoxin ≤0.05 EU/mg).

PA007131: In Vivo Grade Recombinant Human IgG4-S228P L235E P329G Isotype Control Antibody (hIgG4 SPLEPG Isotype Control Mutant)

The recombinant human IgG4-S228P L235E P329G isotype control antibody (hIgG4 SPLEPG isotype control mutant, Fc silenced) was produced in mammalian cells.
Immunogen: N/A.
Clone: 4F17.
Isotype: human IgG4, kappa.
Applications: an isotype-matched negative control for human IgG4 SPLEPG 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 recombinant hIgG4 kappa isotype control and mutants are 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

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 IgG4 S228P kappa antibody), concentration, and conjugation format of the experimental primary antibody. The ideal human isotype control antibody has low or no specific binding to any human sample, and humanized or completely human variable regions in addition to the human IgG constant regions.

Isotype control antibodies are frequently used in IHC and Flow cytometry experiments, and in vivo animal study.

Citations of Syd Labs Human IgG4-S228P Isotype Control Antibody:

1. Discovery and characterization of prolactin neutralizing monoclonal antibodies for the treatment of female-prevalent pain disorders
Stephanie Maciuba,et al.MAbs. 2023.PMCID: PMC10498814
“Prolactin (PRL) has recently been demonstrated to elicit female-selective nociceptor sensitization and increase pain-like behaviors in female animals. Here we report the discovery and characterization of first-in-class, humanized PRL neutralizing monoclonal antibodies (PRL mAbs). We obtained two potent and selective PRL mAbs, PL 200,031 and PL 200,039. PL 200,031 was engineered as human IgG1 whereas PL 200,039 was reformatted as human IgG4. Both mAbs have sub-nanomolar affinity for human PRL (hPRL) and produce concentration-dependent and complete inhibition of hPRL signaling at the hPRL receptor (hPRLR). These two PRL mAbs are selective for hPRL as they do not inhibit other hPRLR agonists such as human growth hormone or placental lactogen. They also cross-react with non-human primate PRL but not with rodent PRL. Further, both mAbs show long clearance half-lives after intravenous administration in FcRn-humanized mice. Consistent with their isotypes, these mAbs only differ in binding affinities to Fcγ receptors, as expected by design. Finally, PL 200,019, the murine parental mAb of PL 200,031 and PL 200,039, fully blocked stress-induced and PRL-dependent pain behaviors in female PRL-humanized mice, thereby providing in vivo preclinical proof-of-efficacy for PRL mAbs in mechanisms relevant to pain in females.”
Tags: activity of Human IgG4-S228P Isotype Control; activity of Human IgG4-S228P Isotype Control mAb

2. Enhanced IL-15-mediated NK cell activation and proliferation by an ADAM17 function-blocking antibody involves CD16A, CD137, and accessory cells
Anders W Matson,et al.J Immunother Cancer. 2024.PMCID: PMC11284835
“Background：Natural killer (NK) cells are being extensively studied as a cell therapy for cancer. These cells are activated by recognition of ligands and antigens on tumor cells. Cytokine therapies, such as IL-15, are also broadly used to stimulate endogenous and adoptively transferred NK cells in patients with cancer. These stimuli activate the membrane protease ADAM17, which cleaves various cell-surface receptors on NK cells as a negative feedback loop to limit their cytolytic function. ADAM17 inhibition can enhance IL-15-mediated NK cell proliferation in vitro and in vivo. In this study, we investigated the underlying mechanism of this process.Methods：Peripheral blood mononuclear cells (PBMCs) or enriched NK cells from human peripheral blood, either unlabeled or labeled with a cell proliferation dye, were cultured for up to 7 days in the presence of rhIL-15±an ADAM17 function-blocking antibody. Different fully human versions of the antibody were generated; Medi-1 (IgG1), Medi-4 (IgG4), Medi-PGLALA, Medi-F(ab′)2, and TAB16 (anti-ADAM17 and anti-CD16 bispecific) to modulate CD16A binding. Flow cytometry was used to assess NK cell proliferation and phenotypic markers, immunoblotting to examine CD16A signaling, and IncuCyte-based live cell imaging to measure NK cell antitumor activity.Results：The ADAM17 function-blocking monoclonal antibody (mAb) Medi-1 markedly increased early NK cell activation by IL-15. By using different engineered versions of the antibody, we demonstrate involvement by CD16A, an activating Fcγ receptor and well-described ADAM17 substrate. Hence, Medi-1 when bound to ADAM17 on NK cells is engaged by CD16A and blocks its shedding, inducing and prolonging its signaling. This process did not promote evident NK cell fratricide or dysfunction. Synergistic signaling by Medi-1 and IL-15 enhanced the upregulation of CD137 on CD16A+ NK cells and augmented their proliferation in the presence of PBMC accessory cells or an anti-CD137 agonistic mAb.Conclusions：Our data reveal for the first time that CD16A and CD137 underpin Medi-1 enhancement of IL-15-driven NK cell activation and proliferation, respectively, with the latter requiring PBMC accessory cells. The use of Medi-1 represents a novel strategy to enhance IL-15-driven NK cell proliferation, and it may be of therapeutic importance by increasing the antitumor activity of NK cells in patients with cancer.”
Tags: human IgG1 recombinant isotype control; Human IgG4-S228P Isotype Control antibody for cancer research

3. DEspRhigh neutrophils are associated with critical illness in COVID-19
Joanne T. deKay,et al.Sci Rep. 2021.PMCID: PMC8599677
“SARS-CoV-2 infection results in a spectrum of outcomes from no symptoms to widely varying degrees of illness to death. A better understanding of the immune response to SARS-CoV-2 infection and subsequent, often excessive, inflammation may inform treatment decisions and reveal opportunities for therapy. We studied immune cell subpopulations and their associations with clinical parameters in a cohort of 26 patients with COVID-19. Following informed consent, we collected blood samples from hospitalized patients with COVID-19 within 72 h of admission. Flow cytometry was used to analyze white blood cell subpopulations. Plasma levels of cytokines and chemokines were measured using ELISA. Neutrophils undergoing neutrophil extracellular traps (NET) formation were evaluated in blood smears. We examined the immunophenotype of patients with COVID-19 in comparison to that of SARS-CoV-2 negative controls. A novel subset of pro-inflammatory neutrophils expressing a high level of dual endothelin-1 and VEGF signal peptide-activated receptor (DEspR) at the cell surface was found to be associated with elevated circulating CCL23, increased NETosis, and critical-severity COVID-19 illness. The potential to target this subpopulation of neutrophils to reduce secondary tissue damage caused by SARS-CoV-2 infection warrants further investigation.”
Tags: in vivo study of Human IgG4-S228P Isotype Control antibody; bulk Human IgG4-S228P Isotype Control

For more references about Human IgG4-S228P Isotype Control Antibody please contact our scientific support team with message@sydlabs.com.

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