Introduction
The NPHS2 gene encodes podocin, a protein crucial for maintaining the glomerular filtration barrier in kidneys. Mutations in this gene lead to steroid-resistant nephrotic syndrome, a condition characterized by proteinuria and edema.
Description
This product consists of a non-glycosylated polypeptide chain of human NPHS2, spanning from amino acid Val125 to Leu383. It's expressed in E. coli and purified to a single band on SDS-PAGE. A 10 amino acid His tag is present at the N-terminus, resulting in a total calculated molecular weight of 30.2kDa.
Physical Appearance
The product appears as a white powder after lyophilization (freeze-drying) and filtration.
Formulation
To ensure stability, the NPHS2 protein was filtered through a 0.4 micrometer filter and subsequently lyophilized in a buffer consisting of 30mM acetate and 1mM dithiothreitol at a pH of 4.
Solubility
To prepare a working solution, it is recommended to reconstitute the lyophilized powder in 0.1M acetate buffer (pH 4.0) to achieve a concentration of approximately 0.5mg/ml. Allow sufficient time for complete dissolution. For use at higher pH values, dilute the stock solution with the appropriate buffer to a concentration of 10µg/ml. It's crucial to note that the solubility of this protein is limited at higher concentrations. For cell culture applications, ensure sterility by filtering the product through a sterile filter before use.
Stability
For long-term storage, store the lyophilized protein at -20°C. Once reconstituted, aliquot the protein to prevent repeated freeze-thaw cycles. While the reconstituted protein can be stored at 4°C for a limited period (approximately two weeks) without noticeable changes, it's recommended to use it promptly.
Purity
Analysis by SDS-PAGE confirms a purity greater than 95.0%.
Synonyms
Nephrosis 2, Idiopathic, Steroid-Resistant (Podocin), PDCN, SRN1, Podocin, NPHS2.
Amino Acid Sequence
MKHHHHHHASVKVVQEYERV IIFRLGHLLP GRAKGPGLFF FLPCLDTYHK VDLRLQTLEI PFHEIVTKDM FIMEIDAICY YRMENASLLL SSLAHVSKAV QFLVQTTMKR LLAHRSLTEI LLERKSIAQD AKVALDSVTC IWGIKVERIE IKDVRLPAGL QHSLAVEAEA QRQAKVRMIA AEAEKAASES LRMAAEILSG TPAAVQLRYL HTLQSLSTEK PSTVVLPLPF DLLNCLSSPS NRTQGSLPFP SPSKPVEPLN PKKKDSPML.