Mouse TNF Alpha/Tumor necrosis factor ELISA Kit EZ-SetTM (DIY Antibody Pairs)

Tumor necrosis factor-alpha (TNF-alpha, or TNF) is secreted by macrophages in response to inflammation, infection and cancer. Human Tumor Necrosis Factor (TNF) and Lymphotoxin (TNF-beta) are cytotoxic proteins which have similar biological activities and share 30% amino acid homology. TNF-alpha is produced by monocytes, which can stimulate endothelial cells to produce the multilineage growth factor granulocyte-macrophage colony-stimulating factor and extend the role of this immunoregulatory protein to the regulation of hematopoiesis in vitro. TNF is a soluble protein that causes damage to tumor cells but has no effect on normal cells. Human TNF has been purified to apparent homogeneity as a 17.3-kilodalton protein from HL-60 leukemia cells and has showed cytotoxic and cytostatic activities against various human tumor cell lines. The human TNF cDNA is 1585 base pairs in length and encodes a protein of 233 amino acids. The mature protein begins at residue 77, leaving a long leader sequence of 76 amino acids. TNF-alpha has been mapped to human chromosome 6.

Catalog number Description Quantity Storage of opened/reconstituted material EZ0527-CA Mouse anti- mouse Tnf monoclonal antibody (Capture Antibody) 500 μl, 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. EZ0527-DA Biotinylated goat anti- mouse Tnf polyclonal antibody (Detection Antibody) 500 μl, 0.15 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK0527-ST Lyophilized recombinant mouse Tnf 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 EZ0527-CA Mouse anti- mouse Tnf monoclonal antibody (Capture Antibody) 500 μl, 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. EZ0527-DA Biotinylated goat anti- mouse Tnf polyclonal antibody (Detection Antibody) 500 μl, 0.15 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK0527-ST Lyophilized recombinant mouse Tnf 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 EZ0527-CA Mouse anti- mouse Tnf monoclonal antibody (Capture Antibody) 500 μl, 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. EZ0527-CA Mouse anti- mouse Tnf monoclonal antibody (Capture Antibody) 500 μl, 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. EZ0527-DA Biotinylated goat anti- mouse Tnf polyclonal antibody (Detection Antibody) 500 μl, 0.15 μg/mL (recommended dilution 1:100) EZ0527-DA Biotinylated goat anti- mouse Tnf polyclonal antibody (Detection Antibody) 500 μl, 0.15 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) EK0527-ST Lyophilized recombinant mouse Tnf 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. EK0527-ST Lyophilized recombinant mouse Tnf 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. 0.051 0.090 0.156 0.281 0.443 0.723 1.284 1.958 Concentration (pg/ml) 0 15.6 31.2 62.5 125 250 500 1000 O.D. 0.051 0.090 0.156 0.281 0.443 0.723 1.284 1.958 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. 0.051 0.090 0.156 0.281 0.443 0.723 1.284 1.958 O.D. 0.051 0.090 0.156 0.281 0.443 0.723 1.284 1.958

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)