Human IgG1-D265A 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, 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.

 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.

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

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.

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