Specificity: LEPTIN
Product Page: Lep antibody (OASA06835)
Antibody Pair Available: OASA06835 Capture OASA06836 Detection
Experiment Type: Leptin measurement

Protocol:
Plasma leptin concentrations were determined with an enzyme immunoassay test available from SpiBio (Montigny le Bretonneux, France). This test is a double-antibody sandwich technique, using biotin-tagged polyclonal anti-rat leptin antibody and streptavidin-horseradish peroxidase tracer. 3,3′5,5′-Tetramethylbenzidine (TMB) reagent forms a yellow compound, which emits at 450 nm. Results are reported to a standard curve drawn under the same conditions with purified rat leptin. Sensitivity of the assay is 50 pg leptin/ml (15-μl sample size).

Summary:
1. Cytochemical controls consisting of samples with primary antibody omitted displayed no labeling. 2. At 30 min, anti-leptin and anti-FITC antibodies combined to protein A-gold revealed the presence of FITC-leptin on enterocyte microvilli, in apical small endocytotic vesicles, at the basolateral membrane interdigitations, and within Golgi cisternae. Junctional complexes appear devoid of labeling. 3. Insertion of recombinant leptin into the duodenal lumen raised plasma leptin concentrations (558 ± 34%) quite rapidly, whereas carrier solution without leptin had no effect.

References:
1: Cammisotto PG, Gingras D, Bendayan M. Transcytosis of gastric leptin through the rat duodenal mucosa. Am J Physiol Gastrointest Liver Physiol. 2007 Oct;293(4):G773-9. Epub 2007 Aug 2. PubMed PMID: 17673543.