FcRn,FCGRT & B2M
Biotinylated Human FcRn Heterodimer Protein (primary amine labeling) (FCM-H8286) is expressed from human 293 cells (HEK293). It contains AA Ala 24 - Ser 297 (FCGRT) & Ile 21 - Met 119 (B2M) (Accession # NP_001129491.1 (FCGRT) & AAH32589 (B2M)). It is the biotinylated form of Human FcRn / FCGRT & B2M Heterodimer Protein (FCM-H5286).
Predicted N-terminus: Ala 24(FCGRT) & Ile 21(B2M)
Biotinylated Human FcRn Heterodimer Protein (primary amine labeling), produced by co-expression of FCGRT and B2M, has a calculated MW of 31.2 kDa (FCGRT) and 13.1 kDa (B2M). Subunit FCGRT is fused with a polyhistidine tag at the C-terminus and subunit Beta-2 microglobulin (B2M) is fused with Strep II-tag at the C-terminus. The reducing (R) protein migrates as 33 kDa (FCGRT) and 13 kDa (B2M) respectively due to glycosylation.
The primary amines in the side chains of lysine residues and the N-terminus of the protein are conjugated with biotins using standard chemical labeling method. A standard biotin reagent (13.5 angstroms) is used in this product.
The biotin to protein ratio is 2.5-4.5 as determined by the HABA assay.
Less than 1.0 EU per μg by the LAL method.
>95% as determined by SDS-PAGE.
Lyophilized from 0.22 μm filtered solution in PBS, pH7.4. Normally trehalose is added as protectant before lyophilization.
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Please see Certificate of Analysis for specific instructions.
For best performance, we strongly recommend you to follow the reconstitution protocol provided in the CoA.
For long term storage, the product should be stored at lyophilized state at -20°C or lower.
Please avoid repeated freeze-thaw cycles.
No activity loss was observed after storage at:
- 4-8°C for 12 months in lyophilized state;
- -70°C for 3 months under sterile conditions after reconstitution.
Biotinylated Human FcRn Heterodimer Protein (primary amine labeling) on SDS-PAGE under reducing (R) condition. The gel was stained overnight with Coomassie Blue. The purity of the protein is greater than 95%.
Immobilized Ipilimumab at 10 μg/mL (100 μL/well) can bind Biotinylated Human FcRn Heterodimer Protein (primary amine labeling) (Cat. No. FCM-H8286) with a linear range of 5-78 ng/mL (QC tested).
Immobilized Biotinylated Human FcRn Heterodimer Protein (primary amine labeling) (Cat. No. FCM-H8286) on SA Biosensor, can bind Herceptin with an affinity constant of 0.49 μM as determined in BLI assay (Fortebio Octet 96) (Routinely tested).
FCGRT & B2M heterodimer protein (FcRn complex) consist of two subunits: p51 (equivalent to FCGRT), and p14 (equivalent to beta-2-microglobulin), and forms an MHC class I-like heterodimer. Fc fragment of IgG, receptor, transporter, alpha (FCGRT) binds to the Fc region of monomeric immunoglobulins gamma and mediates the uptake of IgG from milk. FCGRT possible role in transfer of immunoglobulin G from mother to fetus. Beta-2-microglobulin (B2M) is a component of the class I major histocompatibility complex (MHC) and involved in the presentation of peptide antigens to the immune system.
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