< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
Interference observed with 1 or more available related molecules.
The Quantikine Mouse BAFF/BLyS Immunoassay is a 4.5 hour solid-phase ELISA designed to measure mouse BAFF/BLyS in cell culture supernates, serum, and plasma. It contains NS0-expressed recombinant mouse BAFF/BLyS and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant factor. Results obtained using natural mouse BAFF/BLyS showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring mouse BAFF/BLyS.
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.
The recovery of mouse BAFF/BLyS spiked to three levels throughout the range of the assay was evaluated.
Average % Recovery
Cell Culture Supernates (n=6)
EDTA Plasma (n=6)
Heparin Plasma (n=6)
To assess the linearity of the assay, samples containing mouse BAFF/BLyS in each matrix were diluted with Calibrator Diluent and assayed.
Preparation and Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
BAFF (B-cell activating factor), also known as BLyS, TALL-1, THANK, and TNFSF13B, is a TNF superfamily ligand that plays a critical role in the development and survival of B lineage cells. It can be expressed as a homotrimer or as a heteromer in association with the related protein APRIL. It is produced by many hematopoietic cell types, and a soluble form can be released by activated neutrophils. Both BAFF and APRIL signal through the receptors BCMA and TACI, and BAFF additionally signals through BAFF R. Mice that overexpress BAFF exhibit elevated B cell numbers, increased formation and size of germinal centers, and symptoms of autoimmunity. In addition, BAFF costimulates T cell activation, promotes a Th1 biased immune response, and promotes the expansion of Treg cells.
B cell Activating Factor
Entrez Gene IDs
10673 (Human); 24099 (Mouse); 498666 (Rat);
ApoL related ligand TALL-1; B lymphocyte stimulator; BAFFB-cell activating factor; B-cell-activating factor; BLyS; BLYSB-lymphocyte stimulator; CD257 antigen; CD257; Dendritic cell-derived TNF-like molecule; DTL; TALL1; TALL-1delta BAFF; TALL1Delta4 BAFF; THANK; TNF- and APOL-related leukocyte expressed ligand 1; TNF and ApoL-related leukocyte expressed ligand 1; TNF homolog that activates apoptosis; TNFSF13B; TNFSF20; tumor necrosis factor (ligand) superfamily, member 13b; tumor necrosis factor (ligand) superfamily, member 20; tumor necrosis factor ligand superfamily member 13B; tumor necrosis factor superfamily, member 13b; tumor necrosis factor-like protein ZTNF4; ZTNF4;
Refer to the product for complete assay procedure.
Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
Prepare all reagents, standard dilutions, and samples as directed in the product insert.
Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
80 µL Assay Diluent
Add 80 µL of Assay Diluent to each well.
40 µL Standard, Control, or Sample
Add 40 µL of Standard, Control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours on a horizontal orbital microplate shaker.
Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.
120 µL Conjugate
Add 120 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours on the shaker.
Aspirate and wash 5 times.
120 µL Substrate Solution
Add 120 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.
120 µL Stop Solution
Add 120 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
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