Human BAFF/TNFSF13B PicoKine® Quick ELISA Kit

BAFF was regularly detected by enzyme-linked immunosorbent assay in brain tissue lysates and in normal spinal fluid, and in astrocytes by double fluorescence microscopy. BAFF was localized in astrocytes close to BAFF-R-expressing immune cells. BAFF receptors were strongly expressed in situ in primary central nervous system (CNS) lymphomas.1 The TNF superfamily member B cell-activating factor (BAFF) plays an important role in humoral immunity and in autoimmune diseases, including RA.Local BAFF gene targeting inhibited proinflammatory cytokine expression, suppressed generation of plasma cells and Th17 cells, and markedly ameliorated joint pathology.2 The B cell activating factor BAFF (BlyS/TALL-1/zTNF4) is a tumor necrosis factor (TNF) -related ligand that promotes B cell survival and binds to three receptors (BCMA, TACI, and the recently described BAFF-R) .3 Human BAFF was mapped to chromosome 13q32-34.4 The standard used in this kit is recombinant soluble human BAFF (A134-L295) with the molecular mass of 19.6KDa.

Reproducibility To assay reproducibility, three samples with differing target protein concentrations were assayed using four different lots. Lots Lot 1 (pg/ml) Lot 2 (pg/ml) Lot 3 (pg/ml) Lot 4 (pg/ml) Mean (pg/ml) Standard Deviation CV (%) Sample 1 125 127 151 129 133 9.97 7.5% Sample 2 539 515 538 530 530 21.73 4.1% Sample 3 1336 1087 1217 1225 1216 79.04 6.5% Lots Lot 1 (pg/ml) Lot 2 (pg/ml) Lot 3 (pg/ml) Lot 4 (pg/ml) Mean (pg/ml) Standard Deviation CV (%) Lots Lot 1 (pg/ml) Lot 2 (pg/ml) Lot 3 (pg/ml) Lot 4 (pg/ml) Mean (pg/ml) Standard Deviation CV (%) Sample 1 125 127 151 129 133 9.97 7.5% Sample 1 125 127 151 129 133 9.97 7.5% Sample 2 539 515 538 530 530 21.73 4.1% Sample 2 539 515 538 530 530 21.73 4.1% Sample 3 1336 1087 1217 1225 1216 79.04 6.5% Sample 3 1336 1087 1217 1225 1216 79.04 6.5%

Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 125 539 1336 125 530 1452 Standard deviation 6.12 40.42 78.82 6.00 43.99 110.35 CV (%) 4.9% 7.5% 5.9% 4.8% 8.3% 7.6% Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 125 539 1336 125 530 1452 Standard deviation 6.12 40.42 78.82 6.00 43.99 110.35 CV (%) 4.9% 7.5% 5.9% 4.8% 8.3% 7.6% Intra-Assay Precision Inter-Assay Precision Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 n 16 16 16 24 24 24 Mean (pg/ml) 125 539 1336 125 530 1452 Mean (pg/ml) 125 539 1336 125 530 1452 Standard deviation 6.12 40.42 78.82 6.00 43.99 110.35 Standard deviation 6.12 40.42 78.82 6.00 43.99 110.35 CV (%) 4.9% 7.5% 5.9% 4.8% 8.3% 7.6% CV (%) 4.9% 7.5% 5.9% 4.8% 8.3% 7.6%

Description|Quantity|Volume Anti-tag Pre-coated 96-well Strip Microplate|1|12 strips of 8 wells Standard||2 Antibody A|1|3ml Antibody B|1|3ml Sample Diluent|1|15ml TBS-T Wash Buffer (25x) |1|12ml Color Developing Reagent (TMB) |1|10ml Stop Solution|1|10ml Adhesive Plate Sealers|2|Piece

Concentration (pg/ml) 0 62.5 125 250 500 1,000 2,000 4,000 O.D. 0.018 0.088 0.119 0.223 0.391 0.716 1.262 1.941 Concentration (pg/ml) 0 62.5 125 250 500 1,000 2,000 4,000 O.D. 0.018 0.088 0.119 0.223 0.391 0.716 1.262 1.941 Concentration (pg/ml) 0 62.5 125 250 500 1,000 2,000 4,000 Concentration (pg/ml) 0 62.5 125 250 500 1,000 2,000 4,000 O.D. 0.018 0.088 0.119 0.223 0.391 0.716 1.262 1.941 O.D. 0.018 0.088 0.119 0.223 0.391 0.716 1.262 1.941

Microplate Reader capable of reading absorbance at 450nm. Automated plate washer (optional) Pipettes and pipette tips capable of precisely dispensing 0.5 μl through 1 ml volumes of aqueous solutions. Multichannel pipettes are recommended for large amount of samples. Deionized or distilled water. 500ml graduated cylinders. Test tubes for dilution. Horizontal orbital microplate shaker capable of maintaining a speed of 500 rpm, amplitude 3mm.