TNFRSF1A ELISA Kit (Human) (OKBB00264)

Catalog No: OKBB00264

Size:96 Tests

Human TNFRSF1A ELISA Kit standard curve

Product Info

• Predicted Species Reactivity: Human
• Application: ELISA
• ELISA Kit Detection Method: Colorimetric, OD450 nm
• ELISA Kit Duration: ~ 3 Hours
• ELISA Kit Principle: Aviva's human TNFsRⅠELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for TNFsRⅠ has been precoated onto 96-well plates. Standards (NS0,L30-T211) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for TNFsRⅠ is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The absorbance of yellow is proportional to the human TNFsRⅠ amount of sample captured in plate.
• ELISA Kit Range: 7.8 pg/ml - 500 pg/ml
• ELISA Kit Reproducibility:

  	Intra-Assay Precision			Inter-Assay Precision
  Sample	1	2	3	1	2	3
  Mean (pg/mL)	74	133	261	85	156	278
  St. Dev.	2.52	6.52	14.9	3.91	7.96	18.07
  %CV	3.40	4.90	5.70	4.6	5.10	6.5

• ELISA Kit Component:
  • Lyophilized recombinant human TNFRSF1A standard: 10 ng/tube x 2
  • Anti-human TNFRSF1A Antibody Well Plate: 96 Wells
  • Sample diluent buffer: 30 mL
  • Biotinylated anti-human TNFRSF1A antibody: 130 uL, dilution 1:100
  • Antibody diluent buffer: 12 mL
  • Avidin-Biotin-Peroxidase Complex (ABC): 130 uL, dilution 1:100
  • ABC diluent buffer: 12 mL
  • TMB color developing agent: 10 mL
  • TMB stop solution: 10 mL
• Additional Information: Range: 7.8pg/ml-500pg/ml
  Principle: Aviva’s human TNF s RⅠ ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. Human TNF s RⅠ specific-specific monoclonal antibodies (clone No. 28401.111) were precoated onto 96-well plates. The human specific detection polyclonal antibodies were biotinylated. The test samples and biotinylated detection antibodies were added to the wells subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the human TNF s RⅠ amount of sample captured in plate.
  Notes: 1. To inspect the validity of experiment operation and the appropriateness of sample dilution proportion, pilot
  experiment using standards and a small number of samples is recommended.
  2. The TMB Color Developing agent is colorless and transparent before using, contact us freely if it is not the case.
  3. Before using the Kit, spin tubes and bring down all components to the bottom of tubes.
  4. Duplicate well assay is recommended for both standard and sample testing.
  5. Don’t let 96-well plate dry, for dry plate will inactivate active components on plate.
  6. Don’t reuse tips and tubes to avoid cross contamination.
  7. To avoid to use the reagents from different batches together.
  8. In order to avoid marginal effect of plate incubation due to temperature difference ( reaction may be stronger in the
  marginal wells), it is suggested that the diluted ABC and TMB solution will be pre-warmed in 37C for 30 min before
  using.
  Background: TNF s RⅠ(also known as Solube tumor necrosis factor receptor 1) is a cytokine receptor that binds tumor necrosis factors (TNF). Chan et al. (2000) found that, in contrast, the p60 (TNFRSF1A) and p80 (TNFRSF1B; 191191) TNFA receptors self-assemble through a distinct functional domain in the TNFR extracellular domain, termed the pre-ligand assembly domain (PLAD), in the absence of ligand. Deletion of the PLAD results in monomeric presentation of p60 or p80. Flow cytometric analysis showed that efficient TNFA binding depends on receptor self-assembly. They also found that other members of the TNF receptor superfamily, including the extracellular domains of TRAIL (TNFRSF10A; 603611), CD40 (109535), and FAS (TNFRSF6; 134637), all self-associate but do not interact with heterologous receptors. By Southern blot analysis of human/Chinese hamster somatic cell hybrid DNA, Milatovich et al. (1991, 1991) mapped the TNFR1 gene to 12pter-cen. Derre et al. (1991) found by nonradioactive in situ hybridization that the type 1 receptor (the p55 TNF receptor) is encoded by a gene located on chromosome 12p13.2.
• Reconstitution and Storage: Store at 4C for 6 months, at -20C for 12 months. Avoid multiple freeze-thaw cycles (Shipped with wet ice.)
• Sample Type: cell culture supernates, serum, plasma (heparin, EDTA, citrate) and urine
• Sensitivity: <1 pg/ml
• Predicted Homology Based on Immunogen Sequence: No detectable cross-reactivity with any other cytokine.
• Reference: 1 Chan, F. K.-M., Chun, H. J., Zheng, L., Siegel, R. M., Bui, K. L., Lenardo, M. J. A domain in TNF receptors that
  mediates ligand-independent receptor assembly and signaling. Science 288: 2351-2354, 2000.
  2. Milatovich, A., Song, K., Heller, R. A., Francke, U. The genes for two tumor necrosis factor receptors, TNFR1 and
  TNFR2, map to human chromosomes 12 (12pter-cen) and 1 (1pter-p32). Cytogenet. Cell Genet. 58: 1980-only, 1991.
  3. Derre, J., Kemper, O., Cherif, D., Nophar, Y., Berger, R., Wallach, D. The gene for the type 1 tumor necrosis factor
  receptor (TNF-R1) is localized on band 12p13. Hum. Genet. 87: 231-233, 1991.
• Specificity: Natural and recombinant human TNFsRⅠ
• Immunogen: Expression system for standard: NS0; Immunogen sequence: L30-T211
• Description: For quantitative detection of human TNFsRâ… in cell culture supernatants, serum, plasma( heparin, EDTA, citrate) and urine.

Target Info

• Gene Symbol: TNFRSF1A
• Gene Full Name: tumor necrosis factor receptor superfamily, member 1A
• Alias Symbols: CD120a, FPF, p55, p55 R, p60, p65, TBP1, TBPI, TNF R, TNF R I, TNF R55, TNF-R1, TNF-RI, TNFAR, TNFR I, TNFR1, TNFR55, TNFR60, TNFRI, TNFRSF1A, TNR1A_HUMAN, Tumor necrosis factor alpha receptor, Tumor necrosis factor binding protein 1, Tumor necrosis factor receptor 1, Tumor necrosis factor receptor superfamily member 1A, Tumor necrosis factor receptor superfamily member 1A, membrane form, Tumor necrosis factor receptor type I, Tumor necrosis factor-binding protein 1
• NCBI Gene Id: 7132
• Protein Name: Tumor necrosis factor receptor superfamily member 1A
• Description of Target: TNFsRⅠ(also known as Solube tumor necrosis factor receptor 1) is a cytokine receptor that binds tumor necrosis factors (TNF). In contrast, the p60 (TNFRSF1A) and p80 (TNFRSF1B) TNFA receptors self-assemble through a distinct functional domain in the TNFR extracellular domain, termed the pre-ligand assembly domain (PLAD), in the absence of ligand. Deletion of the PLAD results in monomeric presentation of p60 or p80. Flow cytometric analysis showed that efficient TNFA binding depends on receptor self-assembly. They also found that other members of the TNF receptor superfamily, including the extracellular domains of TRAIL (TNFRSF10A), CD40, and FAS (TNFRSF6), all self-associate but do not interact with heterologous receptors. By Southern blot analysis of human/Chinese hamster somatic cell hybrid DNA, the TNFR1 gene was mapped to 12pter-cen. And by nonradioactive in situ hybridization that the type 1 receptor (the p55 TNF receptor) is encoded by a gene located on chromosome 12p13.2.
• Uniprot ID: P19438
• Protein Accession #: NP_001056.1
• Nucleotide Accession #: NM_001065.3