protein

Human prolactin antagonist

GHP-40

Recombinant human prolactin, del 1-9 G129R mutant consists of one polypeptide chain containing 190 amino and an additional Ala at N-terminus acids and having a molecular mass of ~ 22 kDa, was purified by proprietary chromatographic techniques as described by Oclon et al. PEDS (in press)

E. coli

Lyophylized white powder

The protein was lyophilized from a concentrated (1mg/ml) solution with 0.02-0.03% NaHCO3.

It is recommended to reconstitute the lyophilized hPRL in sterile water or 0.4% NaHCO3 adjusted tp pH 8-9, not less than 100µg/ml, which can then be further diluted to other aqueous solutions, preferably in presence of carrier protein.

Lyophilized del 1-9 G129R hPRL although stable at room temperature for several weeks, should be stored desiccated below -18C. Upon reconstitution at > 0.1 hPRL mg/ml and up to 3 mg/ml and filter sterilization hPRL can be stored at 4C for several weeks.

Greater than 97.0% as determined by: (a) Gel filtration analysis. (b) Analysis by reducing and non-reducing SDS-PAGE gel.

The sequence of the first six N-terminal amino acids was determined and was found to be Ala-Arg-Ser-Gln-Val-Thr

The purified del 1-9 G129R hPRL consists of > 95% monomers as determined by gel-filtration chromatography.

PLR’s del 1-9 G129R hPRL is fully biologically active as evidenced by inhibiting hPRL-induced proliferation of Nb2 cells or Baf3 cells stably transfected with hPRL receptors. It also interacts at 1:1 molar ratio with human prolactin receptor extracellular domain as documented by SEC and SPR (Biacore analysis).

Less than 0.1 ng/µg (IEU/µg) of del 1-9 G129R hPRL

Protein content: Protein quantitation was carried out by UV spectroscopy at 280 nm using the absorbency value of 0.96 as the extinction coefficient for a 0.1% (1mg/ml) solution at pH 8.0. This value is calculated by theDNAman computer analysis program of protein sequences.

This material is offered by PLR for laboratory research