Mouse Leptin, Lep ELISA Kit

Species Reactivity

Mouse

Detection Method

Sandwich ELISA

Intended Use

Mouse Leptin ELISA development kit contains the key components required for the quantitative measurement of natural and/or recombinant Leptin in a sandwich ELISA format within the range of 20-1,250 pg/mL. Using the ELISA protocol described below, this kit provides sufficient reagents to assay Leptin in approximately 1,000 ELISA plate wells.

Contents of Kit

1. PBS: Dilute 10X PBS to 1X PBS, pH 7.2 in sterile water.
2. Wash Buffer: 0.05% Tween-20 in PBS
3. Block Buffer: 1.0% BSA in PBS
4. Diluent: 1.0% BSA in PBS
5. ELISA microplates (Thermo Fisher Cat. # 456529);
6. BSA (Sigma Cat. # A-7030);
7. Stop Solution 2 M Sulfuric Acid (Sigma Cat. # 339741);
8. Dulbecco's PBS [10X] (Gibco BRL Cat. # 14200-075).

Storage

Store the kit at 4°C upon receipt. For more detailed information, please download the following document on our website.

General Description

Leptin (Greek leptos meaning thin) is a 16 kDa protein hormone that plays a key role in regulating energy intake and energy expenditure, including appetite and metabolism. It is one of the most important adipose derived hormones. The Ob(Lep) gene (Ob for obese, Lep for leptin) is located on chromosome 7 in humans.

Reconstitution And Storage

##Capture Antibody: ## 100μg of antigen-affinity purified goat anti-Leptin. Centrifuge vial prior to opening. Reconstitute in 1.0 ml sterile water for a concentration of 100μg/ml. Following reconstitution the Capture antibodies may be stored at 2-8°C for up to 6 months. For long term storage it is recommended to aliquot into working volumes and store at -70°C in a manual defrost freezer.
##Detection Antibody: ##100μg of biotinylated antigen-affinity purified goat anti-Leptin. Centrifuge vial prior to opening. Reconstitute in 1.0 ml sterile water for a concentration of 100μg/ml. Following reconstitution the Detection antibodies may be stored at 2-8°C for up to 6 months. For long term storage it is recommended to aliquot into working volumes and store at -70°C in a manual defrost freezer.
##Mouse Leptin Standard: ##1 μg of recombinant Leptin. Centrifuge vial prior to opening. Reconstitute in 1.0 ml sterile water for a concentration of 1 μg/ml. The Standard may be stored at 2-8°C for one month or aliquoted and stored at -70°C for up to three months in a manual defrost freezer.
##UltraAvidin-HRP Conjugate: ##40μl vial. Upon receipt, UltraAvidin-HRP conjugate should be stored at 2-8°C, DO NOT FREEZE.
##TMB Liquid Substrate: ##Aspirate and wash plate 4 times. Add 100μl of TMB HRPO Microwell Substrate Standard Kinetic One Component "Ready Use" to each well. Incubate at room temperature and monitorcolor development with an ELISA plate reader at 450 nm with wavelength correction set at 540 nm or 570 nm. The reaction may be stopped after 15–20 minutes by adding 100μl of 2 M sulfuric acid to each well.