AdipoGen Life Sciences

Fc (silent) InVivoKine™ Human IgG1 Control

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AG-35B-0018-C05050 µgCHF 340.00

Figure 1: Fc (silent) InVivoKine™ (human) IgG1 Control (#AG-35B-0018) does not bind to the human Fcg Receptor I. Method: Fcg Receptor I (mouse) is coated on an ELISA plate at 1 µg/ml overnight at RT. Fc (silent) InVivoKine™ (human) IgG1 Control, a control Fc InVivoKine™ (human) IgG1 Control (#AG-35B-0015), or a classical Fc (human) IgG1 Control (rec.) (#AG-35B-0007) are added (starting at 4000 ng/ml with a two-fold serial dilution) during one hour and then detected using an anti-human Fc (HRP) antibody. Both control proteins bind to the mouse Fcg Receptor, while the Fc (LALA-PG)-KIH (human) IgG1 Control (rec.) shows no binding as expected with its LALA-PG mutations.

Figure 2: Protein structure of Fc (silent) InVivoKine™ (human) IgG1 Control (Prod. No. AG-35B-0018).

Specifications / Handling

More Information
Product Details
Synonyms	Fc (LALA-PG) KIH (human) IgG1; Immunoglobulin 1 Fragment Constant
Product Type	Protein
Properties
Source/Host	HEK 293 cells. Produced using animal component-free medium.
Sequence	Fc (LALA-PG) (human) IgG1 (Knobs-into-Holes Technology) (see reference: J.B. Ridgway, et al.; Protein Eng. 9, 617 (1996))
Crossreactivity	Human Mouse
Specificity	The Fc contains the mutations LALA-PG that abolish the interaction between the Fc and FcγRs and therefore Fc undesirable effects. The Fc (LALA-PG) is named Fc (silent).
MW	~28kDa (SDS-PAGE)
Purity	≥95% (SDS-PAGE)
Endotoxin Content	<0.01EU/μg purified protein (LAL test).
Concentration	After reconstitution: 1mg/ml
Reconstitution	Reconstitute with 50µl endotoxin-free water.
Accession Number	P01857
Formulation	Lyophilized. Contains PBS.
Other Product Data	Uniprot: P01857
Shipping and Handling
Shipping	BLUE ICE
Short Term Storage	+4°C
Long Term Storage	-20°C
Handling Advice	After reconstitution, prepare aliquots and store at -20°C. Avoid freeze/thaw cycles. Centrifuge lyophilized vial before opening and reconstitution. PBS containing at least 0.1% BSA should be used for further dilutions.
Use/Stability	Stable for at least 6 months after receipt when stored at -20°C. Working aliquots are stable for up to 3 months when stored at -20°C.
Documents
MSDS	Download PDF
Product Specification Sheet
Datasheet	Download PDF

Scientific Background Information

Product Description

Negative control in experiments with recombinant proteins fused to human IgG1 Fc (LALA-PG)-KIH (Knobs-into-Holes) (e.g. Prod. No. AG-40B-0248). This protein is not an appropriate negative control for other standard Fc protein constructs.

The Fc (silent) InVivoKine™ Human IgG1 Control is produced by using two different vectors, one encoding for the Fc (LALA-PG) Knobs sequence with a small linker (synthesizing a protein of 29kDa) and one encoding for the Fc (LALA-PG) Holes sequence (synthesizing a protein of 28kDa). Both vectors transfected into HEK293 cells produce both Fc molecules (Knobs-into-Holes technology; J.B. Ridgway, et al.; Protein Eng. 9, 617 (1996)) required for dimerization and for secretion of the final protein Fc (silent) InVivoKine™ Human IgG1 Control. The LALA-PG mutations inhibit binding to FcγRs and C1q while FcRn binding and Fc stability remain unaffected.

InVivoKines™ are a new generation of recombinant fusion proteins for immunotherapeutic, preclinical and translational in vivo research, developed and manufactured by AdipoGen Life Sciences.

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