Rat FGF21 ELISA Kit EZ-SetTM (DIY Antibody Pairs)

Fibroblast growth factor 21 is a protein that in humans is encoded by the FGF21 gene. By genomic sequence analysis, Nishimura et al. (2000) identified the FGF21 gene within the 5-prime flanking region of the FUT1 gene on chromosome 19. The protein encoded by this gene is a member of the fibroblast growth factor (FGF) family. FGF family members possess broad mitogenic and cell survival activities and are involved in a variety of biological processes including embryonic development, cell growth, morphogenesis, tissue repair, tumor growth and invasion. FGF21 stimulates glucose uptake in adipocytes but not in other cell types. This effect is additive to the activity of insulin. FGF21 treatment of adipocytes is associated with phosphorylation of FRS2, a protein linking FGF receptors to the Ras/MAP kinase pathway. FGF21 injection in ob/ob mice results in an increase in Glut1 in adipose tissue. FGF21 also protects animals from diet-induced obesity when overexpressed in transgenic mice and lowers blood glucose and triglyceride levels when administered to diabetic rodents.

Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 50 155 638 57 172 749 Standard deviation 2.1 6.67 30.62 2.79 11.01 58.42 CV (%) 4.2% 4.3% 4.8% 4.9% 6.4% 7.8% Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 50 155 638 57 172 749 Standard deviation 2.1 6.67 30.62 2.79 11.01 58.42 CV (%) 4.2% 4.3% 4.8% 4.9% 6.4% 7.8% Intra-Assay Precision Inter-Assay Precision Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 n 16 16 16 24 24 24 Mean (pg/ml) 50 155 638 57 172 749 Mean (pg/ml) 50 155 638 57 172 749 Standard deviation 2.1 6.67 30.62 2.79 11.01 58.42 Standard deviation 2.1 6.67 30.62 2.79 11.01 58.42 CV (%) 4.2% 4.3% 4.8% 4.9% 6.4% 7.8% CV (%) 4.2% 4.3% 4.8% 4.9% 6.4% 7.8%

Catalog number Description Quantity Storage of opened/reconstituted material EZ1595-CA Rabbit anti- rat Fgf21 polyclonal 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. EZ1595-DA Biotinylated goat anti- rat Fgf21 polyclonal antibody (Detection Antibody) 500 μl, 2.1 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK1595-ST Lyophilized recombinant rat Fgf21 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 EZ1595-CA Rabbit anti- rat Fgf21 polyclonal 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. EZ1595-DA Biotinylated goat anti- rat Fgf21 polyclonal antibody (Detection Antibody) 500 μl, 2.1 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK1595-ST Lyophilized recombinant rat Fgf21 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 EZ1595-CA Rabbit anti- rat Fgf21 polyclonal 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. EZ1595-CA Rabbit anti- rat Fgf21 polyclonal 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. EZ1595-DA Biotinylated goat anti- rat Fgf21 polyclonal antibody (Detection Antibody) 500 μl, 2.1 μg/mL (recommended dilution 1:100) EZ1595-DA Biotinylated goat anti- rat Fgf21 polyclonal antibody (Detection Antibody) 500 μl, 2.1 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) EK1595-ST Lyophilized recombinant rat Fgf21 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. EK1595-ST Lyophilized recombinant rat Fgf21 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 31.2 62.5 125 250 500 1000 2000 O.D. 0.071 0.144 0.207 0.352 0.586 0.956 1.569 2.251 Concentration (pg/ml) 0 31.2 62.5 125 250 500 1000 2000 O.D. 0.071 0.144 0.207 0.352 0.586 0.956 1.569 2.251 Concentration (pg/ml) 0 31.2 62.5 125 250 500 1000 2000 Concentration (pg/ml) 0 31.2 62.5 125 250 500 1000 2000 O.D. 0.071 0.144 0.207 0.352 0.586 0.956 1.569 2.251 O.D. 0.071 0.144 0.207 0.352 0.586 0.956 1.569 2.251

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)