Rat TNFα PicoKine® Quick ELISA Kit

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. The standard product used in this kit is recombinant rat TNFalpha, consisting of 157 amino acids with the molecular mass of 17KDa.

Lots Lot 1 (pg/ml) Lot 2 (pg/ml) Lot 3 (pg/ml) Lot 4 (pg/ml) Mean (pg/ml) Standard Deviation CV (%) Sample 1 54 46 47 54 50 2.74 5.5% Sample 2 180 136 210 196 181 12.73 7.1% Sample 3 534 602 810 863 702 30.90 4.4% Lots Lot 1 (pg/ml) Lot 2 (pg/ml) Lot 3 (pg/ml) Lot 4 (pg/ml) Mean (pg/ml) Standard Deviation CV (%) Lots Lot 1 (pg/ml) Lot 2 (pg/ml) Lot 3 (pg/ml) Lot 4 (pg/ml) Mean (pg/ml) Standard Deviation CV (%) Sample 1 54 46 47 54 50 2.74 5.5% Sample 1 54 46 47 54 50 2.74 5.5% Sample 2 180 136 210 196 181 12.73 7.1% Sample 2 180 136 210 196 181 12.73 7.1% Sample 3 534 602 810 863 702 30.90 4.4% Sample 3 534 602 810 863 702 30.90 4.4%

Tumor necrosis factor; Cachectin; TNF-alpha; Tumor necrosis factor ligand superfamily member 2; TNF-a; Tumor necrosis factor, membrane form; N-terminal fragment; NTF; Intracellular domain 1; ICD1; Intracellular domain 2; ICD2; C-domain 1; C-domain 2; Tumor necrosis factor, soluble form; Tnf; Tnfa, Tnfsf2

Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 54 180 534 36 245 786 Standard deviation 3.06 11.79 33.05 2.53 10.49 52.11 CV (%) 5.7% 6.6% 6.2% 7.0% 4.3% 6.6% Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 54 180 534 36 245 786 Standard deviation 3.06 11.79 33.05 2.53 10.49 52.11 CV (%) 5.7% 6.6% 6.2% 7.0% 4.3% 6.6% 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) 54 180 534 36 245 786 Mean (pg/ml) 54 180 534 36 245 786 Standard deviation 3.06 11.79 33.05 2.53 10.49 52.11 Standard deviation 3.06 11.79 33.05 2.53 10.49 52.11 CV (%) 5.7% 6.6% 6.2% 7.0% 4.3% 6.6% CV (%) 5.7% 6.6% 6.2% 7.0% 4.3% 6.6%

Description|Quantity|Volume Pre-coated 96-well Strip Microplate|1|12 strips of 8 wells Standard||2 HRP-Linked Antibody|1|6ml Sample Diluent|1|15ml TBS-T Wash Buffer (25x) |1|12ml Color Developing Reagent (TMB) |1|10ml Stop Solution|1|10ml Adhesive Plate Sealers|2|Piece

Concentration (pg/ml) 0 31.2 62.5 125 250 500 1000 2000 O.D. 0.010 0.031 0.062 0.132 0.203 0.555 0.884 1.838 Concentration (pg/ml) 0 31.2 62.5 125 250 500 1000 2000 O.D. 0.010 0.031 0.062 0.132 0.203 0.555 0.884 1.838 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. 0.010 0.031 0.062 0.132 0.203 0.555 0.884 1.838 O.D. 0.010 0.031 0.062 0.132 0.203 0.555 0.884 1.838

Microplate Reader capable of reading absorbance at 450nm. Automated plate washer (optional) Pipettes and pipette tips capable of precisely dispensing 0.5 μl through 1 ml volumes of aqueous solutions. Multichannel pipettes are recommended for large amount of samples. Deionized or distilled water. 500ml graduated cylinders. Test tubes for dilution. Horizontal orbital microplate shaker capable of maintaining a speed of 500 rpm, amplitude 3mm.