Human NGF/NGF beta ELISA Kit EZ-SetTM (DIY Antibody Pairs)

Nerve growth factor (NGF) is a polypeptide involved in the regulation of growth and differentiation of sympathetic and certain sensory neurons. NGF is thought to have a profound effect on the development and maintenance of sympathetic and embryonic sensory neurones. NGF activity isolated from the male mouse submaxillary gland (MSG) consists of three types of subunits, alpha, beta and gamma, which specifically interact to form a 7S, approximately 130,000-molecular weight (Mr) complex. The 7S complex contains two identical 118-amino acid beta-chains, which are solely responsible for the nerve growth-stimulating activity of NGF. NGF, which is expressed by inflammatory cells and effects changes that lead to increased neural responsiveness, could be a pivotal mediator in allergic rhinitis. The standard product used in this kit is human 2.5S NGF, which is a dimmer linking with two polypeptide chains of 120 amino acids.

Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 86.3 201.2 469 93.7 231.6 509.2 Standard deviation 5.6 14.5 34.2 7.1 18.8 39.2 CV (%) 6.5% 7.2% 7.3% 7.6% 8.1% 7.7% Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 86.3 201.2 469 93.7 231.6 509.2 Standard deviation 5.6 14.5 34.2 7.1 18.8 39.2 CV (%) 6.5% 7.2% 7.3% 7.6% 8.1% 7.7% 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) 86.3 201.2 469 93.7 231.6 509.2 Mean (pg/ml) 86.3 201.2 469 93.7 231.6 509.2 Standard deviation 5.6 14.5 34.2 7.1 18.8 39.2 Standard deviation 5.6 14.5 34.2 7.1 18.8 39.2 CV (%) 6.5% 7.2% 7.3% 7.6% 8.1% 7.7% CV (%) 6.5% 7.2% 7.3% 7.6% 8.1% 7.7%

Nerve growth factor is important for the development and maintenance of the sympathetic and sensory nervous systems. Extracellular ligand for the NTRK1 and NGFR receptors, activates cellular signaling cascades through those receptor tyrosine kinase to regulate neuronal proliferation, differentiation and survival. Inhibits metalloproteinase dependent proteolysis of platelet glycoprotein VI (PubMed:20164177) .

Catalog number Description Quantity Storage of opened/reconstituted material EZ0469-CA Rabbit anti- human NGF 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. EZ0469-DA Biotinylated goat anti- human NGF polyclonal antibody (Detection Antibody) 500 μl, 7 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK0469-ST Lyophilized recombinant human NGF 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 EZ0469-CA Rabbit anti- human NGF 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. EZ0469-DA Biotinylated goat anti- human NGF polyclonal antibody (Detection Antibody) 500 μl, 7 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK0469-ST Lyophilized recombinant human NGF 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 EZ0469-CA Rabbit anti- human NGF 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. EZ0469-CA Rabbit anti- human NGF 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. EZ0469-DA Biotinylated goat anti- human NGF polyclonal antibody (Detection Antibody) 500 μl, 7 μg/mL (recommended dilution 1:100) EZ0469-DA Biotinylated goat anti- human NGF polyclonal antibody (Detection Antibody) 500 μl, 7 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) EK0469-ST Lyophilized recombinant human NGF 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. EK0469-ST Lyophilized recombinant human NGF 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 15.6 31.2 62.5 125 250 500 1000 O.D. 0.062 0.091 0.133 0.225 0.403 0.735 1.165 2.059 Concentration (pg/ml) 0 15.6 31.2 62.5 125 250 500 1000 O.D. 0.062 0.091 0.133 0.225 0.403 0.735 1.165 2.059 Concentration (pg/ml) 0 15.6 31.2 62.5 125 250 500 1000 Concentration (pg/ml) 0 15.6 31.2 62.5 125 250 500 1000 O.D. 0.062 0.091 0.133 0.225 0.403 0.735 1.165 2.059 O.D. 0.062 0.091 0.133 0.225 0.403 0.735 1.165 2.059

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)