The sequence of the extracellular domain of human FCGRT (Ala 24-Ser 297) was fused with a C-terminal tag consisting of the AVI tag, TEV protease recognition sequence and a 10-His tag. This was co-transfected with the sequence of human B2M (Ile 21-Met 119) and the resulting FcRn heterodimer was purified.
纯化方法
Nickel and SEC
纯度
> 95 % by SEC-HPLC
内毒素水平
<1.0 EU per mg
Biological Activity Comment
Measured by its binding affinity in a SPR assay on a Biacore 8k instrument. Human FcRn protein, immobilized on a CM5 chip via an anti-His antibody, can bind to anti-HER2 human IgG1 (trastuzumab) with an affinity constant (KD) of 147 nM.
Optimal working dilution should be determined by the investigator.
限制
仅限研究用
状态
Lyophilized
溶解方式
To obtain a final concentration of 1 mg/mL reconstitute 250 μg vials with 250 μL water and 1.0 mg vials with 1.0 mL water. Solubilize for 30 to 60 minutes at room temperature with occasional gentle mixing. Do not vortex.
浓度
1 mg/mL
缓冲液
PBS pH 7.2-7.4 (140 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 1.8 mM KH2PO4)
储存液
Without preservative
储存条件
RT,4 °C,-20 °C,-80 °C
储存方法
Lyophilized proteins are stable at ambient temperature for at least 2 weeks. If the protein is not to be used immediately then the protein should be stored in lyophilized form at -20 °C for up 12 months. Once the protein has been reconstituted we recommend storage at 4 °C for up to one week. For longer term storage of protein in solution we recommend aliquoting into smaller vials to avoid repeated freeze-thaw cycles and storage at -20 or -80 °C for up to 3 months. To avoid surface adsorption loss and inactivation we strongly recommend that the protein should not be aliquoted in less than 10 μg per vial.
FcRn, FCGRT & B2M Background: Neonatal Fc receptor,IgG Fc fragment receptor transporter, also known as the neonatal Fc receptor or more commonly FcRn, consist of two subunits (FCGRT and B2M) and forms an MHC class I-like heterodimer. FcRn binds to the Fc domain of monomeric IgG and mediates the pH dependent recycling of IgG as well as its uptake from milk and transfer from mother to fetus.