Cell Signaling Technology

Product Pathways - Growth Factors/Cytokines

Human BAFF/TNFSF13B (hBAFF) #5233

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Source

Recombinant human BAFF (hBAFF) Ala134-Leu285 (Accession #NP_006564) was expressed in human 293 cells at Cell Signaling Technology.

Molecular Characterization

Recombinant hBAFF contains no "tags" and the nonglycosylated protein has a calculated MW of 15,489. DTT-reduced and non-reduced protein migrate as 16 kDa polypeptides. The expected amino-terminal AVQGP of recombinant hBAFF was verified by amino acid sequencing.

Purity

>98% as determined by SDS-PAGE of 6 μg reduced (+) and non-reduced (-) recombinant hBAFF. All lots are greater than 98% pure.

Bioactivity

The bioactivity of recombinant hBAFF was determined in a cell proliferation assay using mouse splenic B cells. The ED50 of each lot is between 0.5-2 ng/ml.

Coomassie Gel

Coomassie Gel

The purity of recombinant hBAFF was determined by SDS-PAGE of 6 µg reduced (+) and non-reduced (-) recombinant hBAFF and staining overnight with Coomassie Blue.

Bioactivity

Bioactivity

The proliferation of mouse splenic B cells treated with increasing concentrations of hBAFF in the presence of 10 μg/ml goat anti-mouse IgM μ chain was assessed. After 72 hour treatment with hBAFF, cells were incubated with a tetrazolium salt and the OD450-OD650 was determined.

Endotoxin

Less than 0.01 ng endotoxin/1 μg hBAFF.

Formulation

With carrier: Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.2 containing 10 mM DTT and 20 μg BSA per 1 μg hBAFF. Cystines are not required for bioactivity. Carrier free: Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.2 containing 10 mM DTT. Cystines are not required for bioactivity.

Background

BAFF, a member of the TNF superfamily of proteins, is a homotrimeric transmembrane protein, which is cleaved to produce a soluble cytokine (1). BAFF may also further oligomerize into 60-mer structures (1). BAFF is expressed by neutrophils, macrophages, dendritic cells, activated T cells, and epithelial cells (1,2). BAFF plays a key role in B cell development, survival, and activation (1,3,4). BAFF binds to three distinct receptors, BAFF-R, TACI, and BCMA (1). These receptors are differentially expressed during B cell development and among B cell subsets (1,2,4). While BAFF-R and BCMA bind to the homotrimeric form of BAFF, TACI only binds to membrane bound or higher order BAFF structures (1). The BAFF/ BAFF-R interaction activates both canonical and non-canonical NF-κB pathways, PI3K/Akt, and mTOR (2,4). Activation of the noncanonical NF-κB pathway via BAFF-R is negatively regulated by TRAF3 (5). Elevated levels of BAFF may exacerbate many autoimmune disorders, making it an attractive therapeutic target (2).

  1. Mackay, F. and Schneider, P. (2009) Nat Rev Immunol 9, 491-502.
  2. Moisini, I. and Davidson, A. (2009) Clin Exp Immunol 158, 155-63.
  3. Schiemann, B. et al. (2001) Science 293, 2111-4.
  4. Khan, W.N. (2009) J Immunol 183, 3561-7.
  5. Gardam, S. et al. (2008) Immunity 28, 391-401.

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