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Human His6BAFF/TNFSF13B (hHis6 BAFF) #5413
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The purity of recombinant hHis6BAFF was determined by SDS-PAGE of 6 µg reduced (+) and non-reduced (-) recombinant hHis6BAFF and staining overnight with Coomassie Blue.
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The proliferation of mouse splenic B cells treated with increasing concentrations of hHis6BAFF in the presence of 10 μg/ml of goat anti-mouse IgM μ chain was assessed. After 72 hour treatment with hHis6BAFF, cells were incubated with a tetrazolium salt and the OD450-OD650 was determined.
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Western blot analysis of extracts from mouse splenic B cells, untreated or treated with hHis6BAFF for 24 hours, using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb #4370 (upper) and p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb #4695 (lower).
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Source / Purification
Recombinant human His6BAFF (hHis6BAFF) Ala134-Leu285 (Accession #NP_006564) was expressed in human 293 cells at Cell Signaling Technology.
Product Description
>98% as determined by SDS-PAGE of 6 μg reduced (+) and non-reduced (-) recombinant hHis6BAFF. All lots are greater than 98% pure.
Molecular Formula:Recombinant N-terminally His6-tagged hBAFF has a calculated MW of 18,066. DTT-reduced and non-reduced protein migrate as 21 kDa polypeptides. The expected amino terminus of recombinant hHis6BAFF was verified by amino acid sequencing.
Bioactivity:The bioactivity of recombinant hHis6BAFF was determined in a cell proliferation assay using mouse splenic B cells. The ED50 of each lot is between 0.5-2 ng/ml.
Endotoxin:Less than 0.01 ng endotoxin/1 μg hHis6BAFF.
Product Usage Information
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 hHis6BAFF. 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.
Storage: Stable in lyophilized state at 4°C for 1 year after receipt. Sterile stock solutions reconstituted with carrier protein are stable at 4°C for 2 months and at -20°C for 6 months. Avoid repeated freeze-thaw cycles.Maintain sterility. Storage at -20°C should be in a manual defrost freezer.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 signaling (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.
Data Sheets & Documentation
For Research Use Only. Not For Use In Diagnostic Procedures. Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
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