BAFF/BLyS/TNFSF13B Antibody (148725) [Unconjugated]

Recombinant Human BAFF/BLyS/TNFSF13B (Catalog # 2149-BF) stimulates proliferation in mouse B cells in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Human BAFF/BLyS/TNFSF13B (5 ng/mL) is neutralized (green line) by increasing concentrations of Mouse Anti-Human BAFF/BLyS/TNFSF13B Monoclonal Antibody (Catalog # MAB124). At 20 µg/mL, this anti-BAFF/BLyS/TNFSF13B antibody will neutralize >60% of Recombinant Human BAFF/BLyS/TNFSF13B-induced proliferation of mouse B cells.

BAFF-mediated signaling synergizes with LPS for NF-kappa B activation without affecting I kappa B degradation or NF-kappa B nuclear translocation.(A and B) HEK 293 T cells were transfected with an NF-kappa B luciferase construct, reference reporter construct, and BAFF expression vector in combination with CD4-TLR4, death domain (DD) of MyD88, or wild type TRIF-expressing vectors for 24 h. Relative luciferase activity (RLA) was then measured. ***p < 0.001 when compared with the negative control. #p < 0.05, ## < 0.01 and ### < 0.001 (n = 6). (C) THP-1 cells were pre-treated with 1 μg/ml of anti-BAFF mAb or mIgG and then stimulated with 100 ng/ml of LPS for the indicated time periods and total cell lysates were obtained. The Western blot was performed using antibodies specific for phospho-I kappa B-alpha , I kappa B-alpha or beta -actin. (D) Cells were pretreated as in C and stimulated with 1 μg/ml of LPS. After 30 min, the location of p65 was analyzed using immunocytochemistry and the proportion of cells with nuclear p65 was determined (n = 3). Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/28374824), licensed under a CC-BY license. Not internally tested by R&D Systems.

The enhancing effect of mAb is BAFF specific and starts at relatively early time points after LPS stimulation.(A) THP-1 cells were transiently transfected with 10 nM scramble siRNA (NC) or BAFF-specific siRNA. After 24 h, RT-PCR analysis was performed to evaluate the expression levels of BAFF mRNA. (B) Cells were transfected as in (A) and collected at 48 h. BAFF expression levels were then measured by flow cytometry using BAFF specific mAb (empty area) or mIgG (filled area). (C) Cells were transfected as in (A). After 48 h, cells were pre-treated with 1 μg/ml mIgG or anti-BAFF mAb for 30 min and then the cells were stimulated with 100 ng/ml LPS for 24 h. Cultured supernatants were then collected to evaluate levels of secreted IL-8. Values represent IL-8 levels relative to those of the positive control (set to 100%). *p < 0.05 (n = 3). (D) THP-1 cells were pretreated as in (C) and stimulated with 1 μg/ml of LPS. Culture supernatants were collected at the indicated time points and IL-8 levels were measured using ELISA. Control samples were treated with 1 μg/ml anti-BAFF or mIgG for 24 h in the absence of LPS treatment. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/28374824), licensed under a CC-BY license. Not internally tested by R&D Systems.

The PI3K/AKT/CREB pathway contributes to the synergistic activation of NF-kappa B and expression of IL-8.(A) THP-1 cells were pre-treated with anti-BAFF or mIgG (1 μg/ml) for 30 min and then stimulated with 100 ng/ml of LPS for the indicated times. Total cell lysates were tested for the levels of phospho-AKT(Ser473), AKT, and beta -actin by Western blot. (B) THP-1 cells were pre-treated with 5 μM LY294002 (PI3K inhibitor), 10 μM MK2206 (AKT inhibitor) or 0.05% DMSO (VC) for 1 h. The cells were treated with anti-BAFF (1 μg/ml) for 30 min and then stimulated with LPS (100 ng/ml) for 24 h. The levels of secreted IL-8 in culture supernatants were determined by ELISA. **p < 0.01 when compared with corresponding positive control samples stimulated in the absence of inhibitors (n = 6). (C) HEK 293 T cells were co-transfected with an NF-kappa B luciferase construct, reference reporter construct, and BAFF and CD4-TLR4 expressing vectors. After 3 h, the cells were treated with 20 μM LY294002 (PI3K inhibitor), 10 μM MK2206 (AKT inhibitor) or 0.2% DMSO (VC). After 21 h, relative luciferase activities (RLA) were measured. **p < 0.01 when compared with cells co-transfected with BAFF and CD4-TLR4 expressing vectors (n = 6). (D) THP-1 cells were pre-treated with 20 μM LY294002 (PI3K inhibitor) or 0.2% DMSO (VC) for 1 h. The cells were stimulated with LPS (100 ng/ml) for indicated times in the presence of anti-BAFF mAb (1 μg/ml). The phosphorylation levels of AKT and CREB were analyzed by Western blot. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/28374824), licensed under a CC-BY license. Not internally tested by R&D Systems.

BAFF exerts its enhancing effects through activation of ERK and JNK MAPKs.(A) THP-1 cells were stimulated with anti-BAFF (1 μg/ml) mAb for the indicated time periods. The activation status of MAPK was analyzed by Western blot using antibodies specific for normal and phosphorylated form of MAPKs and actin. (B) The cells were pre-treated with 1 μg/ml of anti-BAFF mAb or mIgG for 30 min and then stimulated with 100 ng/ml of LPS for the indicated time periods. The Western blot analysis of MAPKs was performed as in A. (C-F) THP-1 cells were pre-treated with 3.75 μM U0126 (ERK inhibitor), 2 μM SB203580 (p38 inhibitor), 10 μM SP600125 (JNK inhibitor) or 0.02% DMSO (VC) for 30 min. Cells were then treated with 1 μg/ml anti-BAFF for 30 min before stimulation with 100 ng/ml of LPS for 24 h. Culture supernatants were collected and levels of secreted MMP-9 (C), IL-8 (D), TNF-alpha (E) and MCP-1 (F) were analyzed by gelatin zymography and ELISA. *p < 0.05 when compared with corresponding positive control samples stimulated in the absence of inhibitors (n = 3). Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/28374824), licensed under a CC-BY license. Not internally tested by R&D Systems.

BAFF exerts its enhancing effects through activation of ERK and JNK MAPKs.(A) THP-1 cells were stimulated with anti-BAFF (1 μg/ml) mAb for the indicated time periods. The activation status of MAPK was analyzed by Western blot using antibodies specific for normal and phosphorylated form of MAPKs and actin. (B) The cells were pre-treated with 1 μg/ml of anti-BAFF mAb or mIgG for 30 min and then stimulated with 100 ng/ml of LPS for the indicated time periods. The Western blot analysis of MAPKs was performed as in A. (C-F) THP-1 cells were pre-treated with 3.75 μM U0126 (ERK inhibitor), 2 μM SB203580 (p38 inhibitor), 10 μM SP600125 (JNK inhibitor) or 0.02% DMSO (VC) for 30 min. Cells were then treated with 1 μg/ml anti-BAFF for 30 min before stimulation with 100 ng/ml of LPS for 24 h. Culture supernatants were collected and levels of secreted MMP-9 (C), IL-8 (D), TNF-alpha (E) and MCP-1 (F) were analyzed by gelatin zymography and ELISA. *p < 0.05 when compared with corresponding positive control samples stimulated in the absence of inhibitors (n = 3). Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/28374824), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Hu
• Applications: B/N
• Clone: 148725
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: LYOPH

BAFF/BLyS/TNFSF13B Antibody (148725) [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant human BAFF
  Ala134-Leu285
  Accession # Q9Y275
• Specificity: Detects human BAFF in direct ELISAs.
• Source: N/A
• Isotype: IgG2b
• Clonality: Monoclonal
• Host: Mouse
• Gene: TNFSF13B
• Purity Statement: Protein A or G purified from hybridoma culture supernatant
• Endotoxin Note: <0.10 EU per 1 μg of the antibody by the LAL method.

Applications/Dilutions

• Dilutions:
  • Neutralization 20 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.5 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for BAFF/BLyS/TNFSF13B Antibody (148725) [Unconjugated]

• 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

Background

BAFF, also known as B-lymphocyte stimulator (BLyS), TNF- and APOL-related leukocyte expressed ligand 1 (TALL-1), and TNF homolog that activates apoptosis, NFKB and JNK (THANK), is a TNF ligand superfamily member and has been designated TNFSF13B.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for BAFF/BLyS/TNFSF13B Antibody (MAB124)(6)

Publications using MAB124

• H Touil, A Kobert, N Lebeurrier, A Rieger, P Saikali, C Lambert, L Fawaz, CS Moore, A Prat, J Gommerman, JP Antel, Y Itoyama, I Nakashima, A Bar-Or Human central nervous system astrocytes support survival and activation of B cells: implications for MS pathogenesis J Neuroinflammation, 2018-04-19;15(1):114. 2018-04-19 [PMID: 29673365] (Neutralization, Human)
• Endotoxemia contributes to CD27+ memory B-cell apoptosis via enhanced sensitivity to Fas ligation in patients with Cirrhosis Sci Rep, 2016-11-18;6(0):36862. 2016-11-18 [PMID: 27857173] (Neutralization, Human)
• Giacomini E, Severa M, Rizzo F, Mechelli R, Annibali V, Ristori G, Riccieri V, Salvetti M, Coccia E IFN-beta therapy modulates B-cell and monocyte crosstalk via TLR7 in multiple sclerosis patients. Eur J Immunol, 2013-05-28;43(7):1963-72. 2013-05-28 [PMID: 23636665] (Neutralization, Human)
• Benito-Miguel M, Garcia-Carmona Y, Balsa A, Bautista-Caro MB, Arroyo-Villa I, Cobo-Ibanez T, Bonilla-Hernan MG, De Ayala CP, Sanchez-Mateos P, Martin-Mola E, Miranda-Carus ME IL-15 Expression on RA Synovial Fibroblasts Promotes B Cell Survival. PLoS ONE, 2012-07-09;7(7):e40620. 2012-07-09 [PMID: 22792388] (Flow Cytometry, Neutralization, Human)
• Hallock PT, Xu CF, Park TJ Dok-7 regulates neuromuscular synapse formation by recruiting Crk and Crk-L. Genes Dev., 2010-11-01;24(21):2451-61. 11/1/2010 [PMID: 21041412]
• Fu L, Lin-Lee YC, Pham LV, Tamayo A, Yoshimura L, Ford RJ Constitutive NF-kappaB and NFAT activation leads to stimulation of the BLyS survival pathway in aggressive B-cell lymphomas. Blood, 2006-02-23;107(11):4540-8. 2006-02-23 [PMID: 16497967] (ICC, Human)

Bioinformatics

• Gene Symbol: TNFSF13B
• Uniprot:
  • Q9Y275()