FASL ELISA Kit (Human) (OKBB00143)

Catalog No: OKBB00143

Size:96 Tests

Human FASLG 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 FASL ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for FASL has been precoated onto 96-well plates. Standards(CHO, P134-L281) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for FASL 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 FASL amount of sample captured in plate.
• ELISA Kit Range: 15.6 pg/ml - 1,000 pg/ml
• ELISA Kit Reproducibility:

  	Intra-Assay Precision			Inter-Assay Precision
  Sample	1	2	3	1	2	3
  Mean (pg/mL)	108	257	619	112	249	623
  St. Dev.	5.62	12.59	36.52	9.74	22.66	44.86
  %CV	5.20	4.89	5.89	8.69	9.10	7.20

• ELISA Kit Component:
  • Lyophilized recombinant human FASL standard: 10 ng/tube x 2
  • Anti-human FASL Antibody Well Plate: 96 Wells
  • Sample diluent buffer: 30 mL
  • Biotinylated anti-human FASL 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: 15.6pg/ml-1000pg/ml
  Principle: Aviva’s human FASL ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. Human FASL specific-specific monoclonal antibodies 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 FASL 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: Fas ligand (FasL or CD95L) is a type-II transmembrane protein that belongs to the tumor necrosis factor (TNF) family. Its binding with its receptor induces apoptosis. The human FASL gene consists of approximately 8 kb and is split into 4 exons. Fas ligand/receptor interactions play an important role in the regulation of the immune system and the progression of cancer. Fas ligand or FasL is a homotrimeric type II transmembrane protein. It signals through trimerization of FasR, which spans the membrane of the "target" cell. This trimerization usually leads to apoptosis, or cell death. Soluble Fas ligand is generated by cleaving membrane-bound FasL at a conserved cleavage site by the external matrix metalloproteinase MMP-7.
• 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 and plasma (heparin, EDTA, citrate)
• Sensitivity: <2 pg/ml
• Predicted Homology Based on Immunogen Sequence: No detectable cross-reactivity with any other cytokine.
• Reference: 1. Dhein, J.; Walczak, H.; Baumler, C.; Debatin, K.-M.; Krammer, P. H. Autocrine T-cell suicide mediated by APO-1/(Fas/CD95). Nature 373:
  438-441, 1995.
  2. Itoh, N.; Yonehara, S.; Ishii, A.; Yonehara, M.; Mizushima, S.-I.; Sameshima, M.; Hase, A.; Seto, Y.; Nagata, S. The polypeptide encoded
  by the cDNA for human cell surface antigen Fas can mediate apoptosis. Cell 66: 233-243, 1991.
  3. Oehm, A.; Behrmann, I.; Falk, W.; Pawlita, M.; Maier, G.; Klas, C.; Li-Weber, M.; Richards, S.; Dhein, J.; Trauth, B. C.; Ponstingl, H.;
  Krammer, P. H. Purification and molecular cloning of the APO-1 cell surface antigen, a member of the tumor necrosis factor/nerve growth
  factor receptor superfamily: sequence identity with the FAS antigen. J. Biol. Chem. 267: 10709-10715, 1992.
  4. Arscott, P. L.; Stokes, T.; Myc, A.; Giordano, T. J.; Thompson, N. W.; Baker, J. R., Jr. Fas (CD95) expression is up-regulated on papillary
  thyroid carcinoma. J. Clin. Endocr. Metab. 84: 4246-4252, 1999.
  5. Desbarats, J.; Birge, R. B.; Mimouni-Rongy, M.; Weinstein, D. E.; Palerme, J.-S.; Newell, M. K. Fas engagement induces neurite growth
  through ERK activation and p35 upregulation. Nature Cell Biol. 5: 118-125, 2003.
• Specificity: Natural and recombinant human FASL
• Immunogen: Expression system for standard: CHO; Immunogen sequence: P134-L281
• Description: For quantitative detection of human FASL in cell culture supernatants, serum and plasma(heparin, EDTA, citrate).

Target Info

• Gene Symbol: FASLG
• Gene Full Name: Fas ligand
• Alias Symbols: Apoptosis (APO 1) antigen ligand 1, Apoptosis antigen ligand, APT1LG1, APTL, CD178, CD95 L, CD95 ligand, CD95L, CD95L protein, Fas antigen ligand, FASL, FASLG, TNF superfamily member 6, TNFSF6, Tumor necrosis factor (ligand) superfamily member 6, Tumor necrosis factor ligand superfamily member 6
• NCBI Gene Id: 356
• Protein Name: Tumor necrosis factor ligand superfamily member 6
• Description of Target: Fas ligand (FasL or CD95L) is a type-II transmembrane protein that belongs to the tumor necrosis factor (TNF) family. Its binding with its receptor induces apoptosis. The human FASL gene consists of approximately 8 kb and is split into 4 exons. Fas ligand/receptor interactions play an important role in the regulation of the immune system and the progression of cancer. Fas ligand or FasL is a homotrimeric type II transmembrane protein. It signals through trimerization of FasR, which spans the membrane of the "target" cell. This trimerization usually leads to apoptosis, or cell death. Soluble Fas ligand is generated by cleaving membrane-bound FasL at a conserved cleavage site by the external matrix metalloproteinase MMP-7.
• Uniprot ID: P48023
• Protein Accession #: NP_000630.1
• Nucleotide Accession #: NM_000639.2