Mouse CD40/TNFRSF5 PicoKine® Quick ELISA Kit

CD40 is a cell surface receptor that is expressed on the surface of all mature B cells, most mature B-cell malignancies, and some early B-cell acute lymphocytic leukemias, but is not expressed on plasma cells. The CD40 gene is localized to the long arm of chromosome 20, bands q12-q13.2. The protein encoded by this gene is a member of the TNF-receptor superfamily. This receptor has been found to be essential in mediating a broad variety of immune and inflammatory responses including T cell-dependent immunoglobulin class switching, memory B cell development, and germinal center formation.

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 28 31 34 35 32 2.05 6.4% Sample 2 198 188 199 180 191 7.27 3.8% Sample 3 433 412 455 510 453 13.12 2.9% 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 28 31 34 35 32 2.05 6.4% Sample 1 28 31 34 35 32 2.05 6.4% Sample 2 198 188 199 180 191 7.27 3.8% Sample 2 198 188 199 180 191 7.27 3.8% Sample 3 433 412 455 510 453 13.12 2.9% Sample 3 433 412 455 510 453 13.12 2.9%

Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 28 198 433 32 196 412 Standard deviation 1.54 8.91 26.41 1.22 13.52 23.07 CV (%) 5.5% 4.5% 6.1% 3.8% 6.9% 5.6% Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 28 198 433 32 196 412 Standard deviation 1.54 8.91 26.41 1.22 13.52 23.07 CV (%) 5.5% 4.5% 6.1% 3.8% 6.9% 5.6% 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) 28 198 433 32 196 412 Mean (pg/ml) 28 198 433 32 196 412 Standard deviation 1.54 8.91 26.41 1.22 13.52 23.07 Standard deviation 1.54 8.91 26.41 1.22 13.52 23.07 CV (%) 5.5% 4.5% 6.1% 3.8% 6.9% 5.6% CV (%) 5.5% 4.5% 6.1% 3.8% 6.9% 5.6%

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 15.6 31.2 62.5 125 250 500 1,000 O.D. 0.033 0.159 0.249 0.448 0.785 1.405 1.981 2.319 Concentration (pg/ml) 0 15.6 31.2 62.5 125 250 500 1,000 O.D. 0.033 0.159 0.249 0.448 0.785 1.405 1.981 2.319 Concentration (pg/ml) 0 15.6 31.2 62.5 125 250 500 1,000 Concentration (pg/ml) 0 15.6 31.2 62.5 125 250 500 1,000 O.D. 0.033 0.159 0.249 0.448 0.785 1.405 1.981 2.319 O.D. 0.033 0.159 0.249 0.448 0.785 1.405 1.981 2.319

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.