BAFF ELISA Kit (Mouse) (OKBB00299)

Catalog No: OKBB00299

Size:96 Tests

Mouse BAFF ELISA Kit standard curve

Product Info

• Predicted Species Reactivity: Mouse
• Application: ELISA
• ELISA Kit Detection Method: Colorimetric, OD450 nm
• ELISA Kit Duration: ~ 3 Hours
• ELISA Kit Principle: Aviva's mouse BAFF ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from rat specific for BAFF has been precoated onto 96-well plates. Standards(NS0, A127-L309) 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 mouse BAFF amount of sample captured in plate.
• ELISA Kit Range: 46.9 pg/ml - 3,000 pg/ml
• ELISA Kit Reproducibility:

  	Intra-Assay Precision			Inter-Assay Precision
  Sample	1	2	3	1	2	3
  Mean (pg/mL)	139	312	1508	145	354	1397
  St. Dev.	4.9	11.5	58.8	14.4	33.6	124.3
  %CV	3.52	3.68	3.89	9.93	9.49	8.89

• 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: 46.9pg/ml-3000pg/ml
  Principle: Aviva’s mouse BAFF ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. Mouse BAFFspecific-specific polyclonal antibodies were precoated onto 96-well plates. The mouse 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 mouse 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: B-cell activating factor (BAFF) also known as tumor necrosis factor ligand superfamily member 13B is a protein that in humans is encoded by the TNFLSF13B gene.1,2 BAFF is a cytokine that belongs to the tumor necrosis factor (TNF) ligand family. This cytokine is a ligand for receptors TNFRSF13B/TACI, TNFRSF17/BCMA, and TNFRSF13C/BAFFR. This cytokine is expressed in B cell lineage cells, and acts as a potent B cell activator. It has been also shown to play an important role in the proliferation and differentiation of B cells.3All these ligands act as heterotrimers (i.e. three of the same molecule) interacting with heterotrimeric receptors4, although BAFF has been known to be active as either a hetero- or homotrimer.5 The standard used in this kit is recombinant mouse soluble BAFF (A127-L309) with molecular weight 23.2KDa
• 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: <10 pg/ml
• Predicted Homology Based on Immunogen Sequence: No detectable cross-reactivity with any other cytokine
• Reference: 1. Shu HB, Hu WH, Johnson H (May 1999). "TALL-1 is a novel member of the TNF family that is down-regulated by
  mitogens". J. Leukoc. Biol. 65 (5): 680–3.
  2. Schneider P, MacKay F, Steiner V, Hofmann K, Bodmer JL, Holler N, Ambrose C, Lawton P, Bixler S, Acha-Orbea
  H, Valmori D, Romero P, Werner-Favre C, Zubler RH, Browning JL, Tschopp J (June 1999). "BAFF, a novel ligand
  of the tumor necrosis factor family, stimulates B cell growth". J. Exp. Med. 189 (11): 1747–56.
  3. "Entrez Gene: tumor necrosis factor (ligand) superfamily".
  4. Oren,D.A., Li,Y., Volovik,Y., Morris,T.S., Dharia,C., Das,K., Galperina,O., Gentz,R. and Arnold,E. (2002).
  "Structural basis of BLyS receptor recognition". Nat. Struct. Biol. 9 (4): 288–292.
  5. Daridon C, Youinou P, Pers JO (February 2008). "BAFF, APRIL, TWE-PRIL: who's who?". Autoimmun Rev 7 (4):
  267–71.
• Specificity: Natural and recombinant mouse BAFF
• Immunogen: Expression system for standard: NS0; Immunogen sequence: A127-L309
• Description: For quantitative detection of mouse 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: 24099
• Protein Name: Tumor necrosis factor ligand superfamily member 13B
• Description of Target: B-cell activating factor (BAFF) also known as tumor necrosis factor ligand superfamily member 13B is a protein that in humans is encoded by the TNFLSF13B gene.1,2 BAFF is a cytokine that belongs to the tumor necrosis factor (TNF) ligand family. This cytokine is a ligand for receptors TNFRSF13B/TACI, TNFRSF17/BCMA, and TNFRSF13C/BAFFR. This cytokine is expressed in B cell lineage cells, and acts as a potent B cell activator. It has been also shown to play an important role in the proliferation and differentiation of B cells.3All these ligands act as heterotrimers (i.e. three of the same molecule) interacting with heterotrimeric receptors4, although BAFF has been known to be active as either a hetero- or homotrimer.5 The standard used in this kit is recombinant mouse soluble BAFF (A127-L309) with molecular weight 23.2KDa.
• Uniprot ID: Q9WU72
• Protein Accession #: NP_296371.1
• Nucleotide Accession #: NM_033622.1