BAFF ELISA Kit (Human) (OKBB00298)

Catalog No: OKBB00298

Size:96 Tests

Human BAFF 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 BAFF ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for BAFF has been precoated onto 96-well plates. Standards(NS0, A134-L285) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for BAFF 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 BAFF amount of sample captured in plate.
• ELISA Kit Range: 62.5 pg/ml - 4,000 pg/ml
• ELISA Kit Reproducibility:

  	Intra-Assay Precision			Inter-Assay Precision
  Sample	1	2	3	1	2	3
  Mean (pg/mL)	400	950	2108	425	1093	2357
  St. Dev.	27.2	71.25	90.6	39.1	108.2	228.6
  %CV	6.8	7.5	4.29	9.2	9.89	9.69

• ELISA Kit Component:
  • Lyophilized recombinant human E-Cadherin standard: 10 ng/tube x 2
  • Anti-human E-Cadherin Antibody Well Plate: 96 Wells
  • Sample diluent buffer: 30 mL
  • Biotinylated anti-human E-Cadherin 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: 62.5pg/ml-4000pg/ml
  Principle: Aviva’s human BAFF ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. Human BAFF specific-specific polyclonal antibodies were precoated onto 96-well plates. The human specific detection monoclonal 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 BAFF 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: BAFF was regularly detected by enzyme-linked immunosorbent assay in brain tissue lysates and in normal spinal fluid, and in astrocytes by double fluorescence microscopy. BAFF was localized in astrocytes close to BAFF-R-expressing immune cells. BAFF receptors were strongly expressed in situ in primary central nervous system (CNS) lymphomas.1 The TNF superfamily member B cell-activating factor (BAFF) plays an important role in humoral immunity and in autoimmune diseases, including RA.Local BAFF gene targeting inhibited proinflammatory cytokine expression, suppressed generation of plasma cells and Th17 cells, and markedly ameliorated joint pathology.2 The B cell activating factor BAFF (BlyS/TALL-1/zTNF4) is a tumor necrosis factor (TNF)-related ligand that promotes B cell survival and binds to three receptors (BCMA, TACI, and the recently described BAFF-R).3 Human BAFF was mapped to chromosome 13q32-34.4 The standard used in this kit is recombinant soluble human BAFF (A134-L295) with the molecular mass of 19.6KDa.
• 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)
• Sensitivity: <2 pg/ml
• Predicted Homology Based on Immunogen Sequence: No detectable cross-reactivity with any other cytokine
• Reference: 1. Krumbholz, M., Theil, D., Derfuss, T., Rosenwald, A., Schrader, F., Monoranu, C.-M., Kalled, S. L., Hess, D. M.,
  Serafini, B., Aloisi, F., Wekerle, H., Hohlfeld, R., Meinl, E. BAFF is produced by astrocytes and up-regulated in
  multiple sclerosis lesions and primary central nervous system lymphoma. J. Exp. Med. 201: 195-200, 2005.
  2. Lam, Q. L. K., Ko, O. K. H., Zheng, B.-J., Lu, L. Local BAFF gene silencing suppresses Th17-cell generation and
  ameliorates autoimmune arthritis. Proc. Nat. Acad. Sci. 105: 14993-14998, 2008.
  3. Schiemann, B., Gommerman, J. L., Vora, K., Cachero, T. G., Shulga-Morskaya, S., Dobles, M., Frew, E., Scott, M.
  L. An essential role for BAFF in the normal development of B cells through a BCMA-independent pathway. Science
  293: 2111-2114, 2001.
  4. Schneider, P., MacKay, F., Steiner, V., Hofmann, K., Bodmer, J. L., Holler, N., Ambrose, C., Lawton, P., Bixler, S.,Acha-Orbea, H., Valmori, D., Romero, P., Werner-Favre, C., Zubler, R. H., Browning, J. L., Tschopp, J. BAFF, a novel ligand of the tumor necrosis factor family, stimulates B cell growth. J. Exp. Med. 189: 1747-1756, 1999.
• Specificity: Natural and recombinant human BAFF
• Immunogen: Expression system for standard: NS0; Immunogen sequence: A134-L285
• Description: For quantitative detection of human BAFF in cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA).

Target Info

• Gene Symbol: TNFSF13B
• Gene Full Name: tumor necrosis factor (ligand) superfamily, member 13b
• Alias Symbols: B lymphocyte stimulator, B-cell-activating factor, BAFF, BLyS, CD257, Dendritic cell-derived TNF-like molecule, soluble form, TALL-1, TN13B_HUMAN, TNF- and APOL-related leukocyte expressed ligand 1, TNFSF13B, Tumor necrosis factor ligand superfamily member 13b
• NCBI Gene Id: 10673
• Protein Name: Tumor necrosis factor ligand superfamily member 13B
• Description of Target: BAFF was regularly detected by enzyme-linked immunosorbent assay in brain tissue lysates and in normal spinal fluid, and in astrocytes by double fluorescence microscopy. BAFF was localized in astrocytes close to BAFF-R-expressing immune cells. BAFF receptors were strongly expressed in situ in primary central nervous system (CNS) lymphomas.1 The TNF superfamily member B cell-activating factor (BAFF) plays an important role in humoral immunity and in autoimmune diseases, including RA.Local BAFF gene targeting inhibited proinflammatory cytokine expression, suppressed generation of plasma cells and Th17 cells, and markedly ameliorated joint pathology.2 The B cell activating factor BAFF (BlyS/TALL-1/zTNF4) is a tumor necrosis factor (TNF)-related ligand that promotes B cell survival and binds to three receptors (BCMA, TACI, and the recently described BAFF-R).3 Human BAFF was mapped to chromosome 13q32-34.4 The standard used in this kit is recombinant soluble human BAFF (A134-L295) with the molecular mass of 19.6KDa.
• Uniprot ID: Q9Y275
• Protein Accession #: NP_001139117.1
• Nucleotide Accession #: NM_001145645.2