TNF-a ELISA Kit (Human) : 96 Wells (OKAG00066)

Catalog No: OKAG00066 (Formerly GWB-SKR066)

Size:12x8WELLS

TNF-Alpha ELISA Kit (Human) (OKAG00066) Standard Curve

Product Info

• Predicted Species Reactivity: Homo sapiens, Human
• Product Format: 1 x 96-Well Plate or 12 x 8-Well Strips
• Application: ELISA-Sandwich
• ELISA Kit Detection Method: Colorimetric 450 nm
• ELISA Kit Duration: 4.5 hours
• ELISA Kit Principle: The Aviva Human TNF-alpha ELISA Kit contains the components necessary for quantitative determination of natural or recombinant hTNF-alpha concentrations within any experimental sample including cell lysates, serum and plasma. This particular immunoassay utilizes the quantitative technique of a "Sandwich" Enzyme-Linked Immunosorbent Assay (ELISA) where the target protein (antigen) is bound in a "sandwich" format by the primary capture antibodies coated to each well-bottom and the secondary detection antibodies added subsequently by the investigator. The capture antibodies coated to the bottom of each well are specific for a particular epitope on the Human TNF-alpha cytokine while the user-added detection antibodies bind to epitopes on the captured target protein. Amid each step of the procedure, a series of wash steps must be performed to ensure the elimination of non-specific binding between proteins to other proteins or to the solid phase. After incubation and "sandwiching" of the target antigen, a peroxidase enzyme is conjugated to the constant heavy chain of the secondary antibody (either covalently or via Avidin/Streptavidin-Biotin interactions), allowing for a colorimetric reaction to ensue upon substrate addition. When the substrate TMB (3, 3’, 5, 5’-Tetramethylbenzidine) is added, the reaction catalyzed by peroxidase yields a blue color that is representative of the antigen concentration. Upon sufficient color development, the reaction can be terminated through addition of Stop Solution (2 N Sulfuric Acid) where the color of the solution will turn yellow. The absorbance of each well can then be read by a spectrophotometer, allowing for generation of a standard curve and subsequent determination of protein concentration.
• ELISA Kit Range: 16-2000 pg/ml
• ELISA Kit Component:
  • Capture Antibody Coated Microplate: 96 Wells (12 x 8)
  • Ready-to-Use Streptavidin-HRP: 12 mL
  • Ready-to-Use Substrate: 12 mL
  • Stop Solution: 12 mL
  • Wash Buffer (15x): 50 mL
  • Protein Standard Diluent: 12 mL
  • Sample Diluent: 12 mL
  • Detection Antibody Diluent: 12 mL
  • Biotin-Labeled Detection Antibody: Lyophilized
  • Protein Standard: Lyophilized (100 ng)
• Reconstitution and Storage: 2°C to 8°C
• Sample Type: Cell lysates, sera and plasma
• Sensitivity: 16 pg/mL
• Specificity: The Human TNF-α ELISA is capable of recognizing both recombinant and naturally produced Human TNF-α proteins. The antigens listed below were tested at 50 ng/ml and exhibited less than 1% cross reactivity.
  • Rat: TNF-α
  The antigens listed below were tested at 50 ng/ml and did not exhibit significant cross reactivity or interference.
  • Human: 4-1BBL, 4-1BBR, BAFF, CD40L, sFasL, sRANKL, sTNF-RI, sTNF-RII, TNF-β, sTRAIL, sTRAIL-RI, TWEAK, TWEAK-R, VEGF
• Assay Info: Quantitative Colorimentric Sandwich ELISA

Target Info

• Gene Symbol: TNF
• Gene Full Name: tumor necrosis factor
• Alias Symbols: APC1 protein;Cachectin;DIF;TNF, macrophage-derived;TNF, monocyte-derived;TNF-a;TNFA;TNF-alpha;TNFSF2;TNLG1F;tumor necrosis factor;tumor necrosis factor ligand 1F;tumor necrosis factor ligand superfamily member 2;tumor necrosis factor-alpha.
• NCBI Gene Id: 7124
• Protein Name: P01375
• Description of Target: Tumor necrosis factor
• Uniprot ID: https://www.uniprot.org/uniprot/P01375
• Protein Accession #: NP_000585.2
• Nucleotide Accession #: NM_000594.3