Leptin, Pegylated

Mono-pegylated (with 20 kDa PEG) recombinant human leptin is an one polypeptide chain containing 146 amino, an additional Ala at N-terminus and one molecule of PEG 20 kDa at its N-terminus, having an expected molecular mass of ~ 35.6 kDa as determined by MS. However due to enlarged hydrodymanic volume recombinant mouse pegylated leptin runs on the SDS-PAGE as 48 kDa protein and in gel-filtration on Superdex 200 as over 100 kDa protein. Recombinant human pegylated leptin half-life in circulation after SC injection was over 20 hours. Recombinant human leptin was purified by proprietary chromatographic techniques according to Salomon et al (2006) Protein Expression and Purification 47, 128–136 and then pegylated.

Product Specifications

Synonyms

Lep; ob; obese

NCBI Gene ID

3952

UniProt

P41159

Accession Number

NP_000221.1

Accession Number mRNA

NM_000230

Reactivity

Human

Cross Reactivity

Human

Label

PEG

Sequence

AVPIQKVQDDTKTLIKTIVTRINDISHTQSVSSKQKVTGLDFIPGLHPILTLSKMDQTLAVYQQILTSMPSRNVIQISNDLENLRDLLHVLAFSKSCHLPWASGLETLDSLGGVLEASGYSTEVVALSRLQGSLQDMLWQLDLSPGC

Assay Protocol

It is recommended to reconstitute the lyophilized recombinant human pegylated leptin in sterile water or sterile 0.4% NaHCO3 adjusted to pH 8-9, not less than 100µg/ml, which can then be further diluted with other aqueous solutions.

Endotoxin

< 0.1 ng/µg of protein (< 1EU/µg)

Purity

> 98.0% as determined by Gel filtration and SDS-PAGE gel.

Bioactivity

Recombinant mouse pegylated leptin is capable of stimulatng proliferation of BAF/3 cells stably transfected with the long form of human leptin receptor. Its in vitro activity is 5-7 fold lower than the non-pegylated recombinant human leptin but in vivo it has profound weight reducing effect (as compared to the non-pegylated recombinant human leptin), resulting mainly from reduced food intake.