Mouse BAFF/BLyS/TNFSF13B Quantikine ELISA Kit

  (22 citations)     
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Product Details
Assay Procedure
Citations (22)
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  • Assay Type
    Solid Phase Sandwich ELISA
  • Format
    96-well strip plate
  • Assay Length
    4.5 hours
  • Sample Type & Volume Required Per Well
    Cell Culture Supernates (40 uL), Serum (17 uL), EDTA Plasma (17 uL), Heparin Plasma (17 uL)
  • Sensitivity
    7.8 pg/mL
  • Assay Range
    46.9 - 3,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
  • Specificity
    Natural and recombinant mouse BAFF/BLyS
  • Cross-reactivity
    < 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
  • Interference
    Interference observed with 1 or more available related molecules.
Product Summary
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.

Precision
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
Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma
Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 50 64 56
Mean 125 288 1315 127 325 1262
Standard Deviation 3.6 7 30.3 16 27 136
CV% 2.9 2.4 2.3 12.6 8.3 10.8

Recovery

The recovery of mouse BAFF/BLyS spiked to three levels throughout the range of the assay was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Supernates (n=6) 103 90-108
EDTA Plasma (n=6) 96 85-103
Heparin Plasma (n=6) 94 85-100
Serum (n=6) 98 86-115
Linearity
To assess the linearity of the assay, samples containing mouse BAFF/BLyS in each matrix were diluted with Calibrator Diluent and assayed.
 BAFF/BLyS/TNFSF13B [HRP]
Product Datasheets

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Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: BAFF/BLyS/TNFSF13B
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.
  • Long Name:
    B cell Activating Factor
  • Entrez Gene IDs:
    10673 (Human); 24099 (Mouse); 498666 (Rat)
  • Alternate Names:
    ApoL related ligand TALL-1; B lymphocyte stimulator; BAFF; 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
Related Research Areas
Assay Procedure
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.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

  3. 80 µL Assay Diluent
  4.   Add 80 µL of Assay Diluent to each well.

  5. 40 µL Standard, Control, or Sample
  6.   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.
  7.   Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.

  8. 120 µL Conjugate
  9.   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.
  10.   Aspirate and wash 5 times.

  11. 120 µL Substrate Solution
  12.   Add 120 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.

  13. 120 µL Stop Solution
  14.   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.
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

22 Citations: Showing 1 - 10
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Species
Sample Type
  1. Testosterone is an endogenous regulator of BAFF and splenic B cell number
    Authors: AS Wilhelmson, M Lantero Ro, A Stubelius, P Fogelstran, I Johansson, MB Buechler, S Lianoglou, VN Kapoor, ME Johansson, JB Fagman, A Duhlin, P Tripathi, A Camponesch, BT Porse, AG Rolink, H Nissbrandt, SJ Turley, H Carlsten, IL Mårtensson, MCI Karlsson, Å Tivesten
    Nat Commun, 2018;9(1):2067.
    Species: Mouse
    Sample Type: Serum
  2. Expression of the B cell differentiation factor BAFF and chemokine CXCL13 in a murine model of Respiratory Syncytial Virus infection
    Authors: W Alturaiki, AJ McFarlane, K Rose, R Corkhill, PS McNamara, J Schwarze, BF Flanagan
    Cytokine, 2018;0(0):.
    Species: Mouse
    Sample Type: Tissue Homogenates
  3. B Cell Homeostasis and Functional Properties Are Altered in an Hypochlorous Acid-Induced Murine Model of Systemic Sclerosis
    Authors: S Sanges, M Jendoubi, N Kavian, C Hauspie, S Speca, JC Crave, T Guerrier, G Lefèvre, V Sobanski, A Savina, E Hachulla, PY Hatron, M Labalette, F Batteux, S Dubucquoi, D Launay
    Front Immunol, 2017;8(0):53.
    Species: Mouse
    Sample Type: Serum
  4. Irgm1 coordinately regulates autoimmunity and host defense at select mucosal surfaces
    Authors: KM Azzam, JH Madenspach, DW Cain, L Lai, KM Gowdy, P Rai, K Janardhan, N Clayton, W Cunningham, H Jensen, PS Patel, JF Kearney, GA Taylor, MB Fessler
    JCI Insight, 2017;2(16):.
    Species: Mouse
    Sample Type: Tissue Homogenates
  5. Elevated GDF-15 contributes to pulmonary inflammation upon cigarette smoke exposure
    Authors: FM Verhamme, LJ Seys, EG De Smet, S Provoost, W Janssens, D Elewaut, GF Joos, GG Brusselle, KR Bracke
    Mucosal Immunol, 2017;0(0):.
    Species: Mouse
    Sample Type: Tissue Homogenates
  6. The Deubiquitinase OTULIN Is an Essential Negative Regulator of Inflammation and Autoimmunity
    Cell, 2016;0(0):.
    Species: Mouse
    Sample Type: Serum
  7. BANK1 Regulates IgG Production in a Lupus Model by Controlling TLR7-Dependent STAT1 Activation
    PLoS ONE, 2016;11(5):e0156302.
    Species: Mouse
    Sample Type: Serum
  8. MRL Strains Have a BAFFR Mutation without Functional Consequence
    PLoS ONE, 2016;11(5):e0154518.
    Species: Mouse
    Sample Type: Serum
  9. Nfkb1 activation by the E26 transformation-specific transcription factors PU.1 and Spi-B promotes Toll-like receptor-mediated splenic B cell proliferation.
    Authors: Li S, Abbas A, Solomon L, Groux G, DeKoter R
    Mol Cell Biol, 2015;35(9):1619-32.
    Species: Mouse
    Sample Type: Serum
  10. Impaired B cell receptor signaling is responsible for reduced TACI expression and function in X-linked immunodeficient mice.
    Authors: Uslu K, Coleman A, Allman W, Katsenelson N, Bram R, Alugupalli K, Akkoyunlu M
    J Immunol, 2014;192(8):3582-95.
    Species: Mouse
    Sample Type: Serum
  11. Oral administration of Lactobacillus plantarum strain AYA enhances IgA secretion and provides survival protection against influenza virus infection in mice.
    Authors: Kikuchi, Yosuke, Kunitoh-Asari, Ayami, Hayakawa, Katsuyuk, Imai, Shinjiro, Kasuya, Kenji, Abe, Kimio, Adachi, Yu, Fukudome, Shin-Ich, Takahashi, Yoshimas, Hachimura, Satoshi
    PLoS ONE, 2014;9(1):e86416.
    Species: Mouse
    Sample Type: Cell Culture Supernates
  12. The B lymphocyte differentiation factor (BAFF) is expressed in the airways of children with CF and in lungs of mice infected with Pseudomonas aeruginosa.
    Authors: Neill, Daniel R, Saint, Gemma L, Bricio-Moreno, Laura, Fothergill, Joanne L, Southern, Kevin W, Winstanley, Craig, Christmas, Stephen, Slupsky, Joseph R, McNamara, Paul S, Kadioglu, Aras, Flanagan, Brian F
    PLoS ONE, 2014;9(5):e95892.
    Species: Mouse
    Sample Type: Tissue Homogenates
  13. The development and survival but not function of follicular B cells is dependent on IL-7Ralpha Tyr449 signaling.
    Authors: Patton D, Plumb A, Redpath S, Osborne L, Perona-Wright G, Abraham N
    PLoS ONE, 2014;9(2):e88771.
    Species: Mouse
    Sample Type: Serum
  14. Role of Lactobacillus pentosus Strain b240 and the Toll-like receptor 2 axis in Peyer's patch dendritic cell-mediated immunoglobulin A enhancement.
    Authors: Kotani Y, Kunisawa J, Suzuki Y, Sato I, Saito T, Toba M, Kohda N, Kiyono H
    PLoS ONE, 2014;9(3):e91857.
    Species: Mouse
    Sample Type: Cell Culture Supernates
  15. Recombination-activating gene 1 (Rag1)-deficient mice with severe combined immunodeficiency treated with lentiviral gene therapy demonstrate autoimmune Omenn-like syndrome.
    Authors: van Til N, Sarwari R, Visser T, Hauer J, Lagresle-Peyrou C, van der Velden G, Malshetty V, Cortes P, Jollet A, Danos O, Cassani B, Zhang F, Thrasher A, Fontana E, Poliani P, Cavazzana M, Verstegen M, Villa A, Wagemaker G
    J Allergy Clin Immunol, 2014;133(4):1116-23.
    Species: Mouse
    Sample Type: Plasma
  16. Lack of T cells in Act1-deficient mice results in elevated IgM-specific autoantibodies but reduced lupus-like disease.
    Authors: Johnson AC, Davison LM, Giltiay NV, Vareechon C, Li X, Jorgensen TN
    Eur. J. Immunol., 2012;42(7):1695-705.
    Species: Mouse
    Sample Type: Serum
  17. Mice overexpressing BAFF develop a commensal flora-dependent, IgA-associated nephropathy.
    Authors: McCarthy DD, Kujawa J, Wilson C
    J. Clin. Invest., 2011;121(10):3991-4002.
    Species: Mouse
    Sample Type: Serum
  18. Secreted IgM enhances B cell receptor signaling and promotes splenic but impairs peritoneal B cell survival.
    Authors: Notley CA, Baker N, Ehrenstein MR
    J. Immunol., 2010;184(7):3386-93.
    Species: Mouse
    Sample Type: Serum
  19. The adaptor molecule Act1 regulates BAFF responsiveness and self-reactive B cell selection during transitional B cell maturation.
    Authors: Giltiay NV, Lu Y, Allman D, Jorgensen TN, Li X
    J. Immunol., 2010;185(1):99-109.
    Species: Mouse
    Sample Type: Serum
  20. Emu-BCL10 mice exhibit constitutive activation of both canonical and noncanonical NF-kappaB pathways generating marginal zone (MZ) B-cell expansion as a precursor to splenic MZ lymphoma.
    Authors: Li Z, Wang H, Xue L, Shin DM, Roopenian D, Xu W, Qi CF, Sangster MY, Orihuela CJ, Tuomanen E, Rehg JE, Cui X, Zhang Q, Morse HC, Morris SW
    Blood, 2009;114(19):4158-68.
    Species: Mouse
    Sample Type: Serum
  21. Prevention of murine antiphospholipid syndrome by BAFF blockade.
    Authors: Kahn P, Ramanujam M, Bethunaickan R, Huang W, Tao H, Madaio MP, Factor SM, Davidson A
    Arthritis Rheum., 2008;58(9):2824-34.
    Species: Mouse
    Sample Type: Serum
  22. Inhibition of hedgehog signaling for the treatment of murine sclerodermatous chronic graft-versus-host disease.
    Blood, 0;120(14):2909-17.
    Species: Mouse
    Sample Type: Serum
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