Human Leptin ELISA Kit EZ-SetTM (DIY Antibody Pairs)

Leptin (or obese, OB) is a circulating hormone that is expressed abundantly and specifically in the adipose tissue. Human ob protein is a 166-amino acid polypeptide with a putative signal sequence and is 84 and 83% homologous to the mouse and rat ob proteins, respectively. Northern blot analysis reveals that OB RNA is present at a high level in adipose tissue but at much lower levels in placenta and heart. Indeed, Leptin is first described as an adipocyte-derived signalling factor, which, after interaction with its receptors, induces a complex response including control of bodyweight and energy expenditure. Leptin seems in addition to its role in metabolic control to have important roles in reproduction and neuroendocrine signalling. Leptin is a powerful inhibitor of bone formation in vivo. Comparative mapping of mouse and human DNA indicates that the ob gene is located within a region of mouse chromosome 6 that is homologous to a portion of human chromosome 7q. The standard product used in this kit is recombinant human Leptin.

Catalog number Description Quantity Storage of opened/reconstituted material EZ0437-CA Mouse anti- human LEP monoclonal antibody (Capture Antibody) 500 μl, 0.4 mg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ0437-DA Biotinylated goat anti- human LEP polyclonal antibody (Detection Antibody) 500 μl, 8.8 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK0437-ST Lyophilized recombinant human LEP standard 10 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. Catalog number Description Quantity Storage of opened/reconstituted material EZ0437-CA Mouse anti- human LEP monoclonal antibody (Capture Antibody) 500 μl, 0.4 mg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ0437-DA Biotinylated goat anti- human LEP polyclonal antibody (Detection Antibody) 500 μl, 8.8 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK0437-ST Lyophilized recombinant human LEP standard 10 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. Catalog number Description Quantity Storage of opened/reconstituted material Catalog number Description Quantity Storage of opened/reconstituted material EZ0437-CA Mouse anti- human LEP monoclonal antibody (Capture Antibody) 500 μl, 0.4 mg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ0437-CA Mouse anti- human LEP monoclonal antibody (Capture Antibody) 500 μl, 0.4 mg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ0437-DA Biotinylated goat anti- human LEP polyclonal antibody (Detection Antibody) 500 μl, 8.8 μg/mL (recommended dilution 1:100) EZ0437-DA Biotinylated goat anti- human LEP polyclonal antibody (Detection Antibody) 500 μl, 8.8 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK0437-ST Lyophilized recombinant human LEP standard 10 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. EK0437-ST Lyophilized recombinant human LEP standard 10 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit.

Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles (Ships with gel ice, can store for up to 3 days in room temperature. Freeze upon receiving.)

Concentration (pg/ml) 0 62.5 125 250 500 1000 2000 4000 O.D. 0.002 0.036 0.080 0.191 0.505 0.923 1.801 2.461 Concentration (pg/ml) 0 62.5 125 250 500 1000 2000 4000 O.D. 0.002 0.036 0.080 0.191 0.505 0.923 1.801 2.461 Concentration (pg/ml) 0 62.5 125 250 500 1000 2000 4000 Concentration (pg/ml) 0 62.5 125 250 500 1000 2000 4000 O.D. 0.002 0.036 0.080 0.191 0.505 0.923 1.801 2.461 O.D. 0.002 0.036 0.080 0.191 0.505 0.923 1.801 2.461

1. Microplate reader in standard size. 2. Automated plate washer. 3. Incubator. 4. Adjustable pipettes and pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection. 5. Clean tubes and Eppendorf tubes. 6. 96 well microplate (Cat# AR1100) 7. Plate Sealers. 8. Capture Antibody Diluent: PBS. 9. Reagent Diluent: 1% BSA in PBS, pH 7.2-7.4, 0.2 μm filtered. 10. Substrate Solution: Tetramethylbenzidine (Cat# AR1104) 11. Stop Solution: 2 N H2SO4 (Cat# AR1105) 12. Wash Buffer (PBS and PBS-T) . Recipes: PBS: 8g NaCl, 0.2g KCl, 1.15g Na2HPO4, 0.2g KH2PO4, adjust the total volume to 1 L with distilled water, pH 7.2-7.4, 0.2 μm filtered. PBS-T: 0.1% Tween® 20 in PBS, pH 7.2-7.4. *Item 6 – 12 are included in the EZ Set Accessory Kit (EZA001)