The mean CD40 concentration was determined to be 23,443.1 pg/mL in mouse spleen tissue extract based on a 3.0 mg/mL extract load, 390.2 pg/mL in mouse heart tissue extract based on a 11.2 mg/mL extract load, 216.2 pg/mL in mouse kidney tissue extract based on a 1.5 mg/mL extract load, 3,196.8 pg/mL in mouse liver tissue extract based on a 3.0 mg/mL extract load and 9,508.1 pg/mL in mouse lung tissue extract based on a 3.0 mg/mL extract load.

Mouse spleen tissue, mouse kidney tissue and mouse lung tissue was cultured for 3 days in RPMI supplemented with 10% fetal bovine serum, 5 μM β-mercaptoethanol, 2 mM L-glutamine, 100 U/mL penicillin, 100 μg/mL streptomycin sulfate, and stimulated with 1 μg/mL LPS. The mean CD40 concentration was determined to be 5.7 pg/mL, 37.2 pg/mL and 8.3 pg/mL in unstimulated mouse spleen tissue, mouse kidney tissue and mouse lung tissue supernatant, 6.2 pg/mL, 26.2 pg/mL and 4.5 pg/mL in LPS stimulated mouse spleen tissue, mouse kidney tissue and mouse lung tissue supernatant.

Serum of sixteen mice was measured. The mouse CD40 concentration of detected samples was determined to be 122.5 pg/mL with a range of 10.4 - 236.8 pg/mL.

Sandwich ELISA standard curve of MP00896-1, Mouse CD40 Recombinant Matched Antibody Pair - PBS only. 83972-2-PBS was coated to a plate as the capture antibody and incubated with serial dilutions of standard Eg1118. 83972-1-PBS was HRP conjugated as the detection antibody. Range: 15.6-1000 pg/mL

Cytometric bead array standard curve of MP00896-1, MOUSE CD40 Recombinant Matched Antibody Pair, PBS Only. Capture antibody: 83972-2-PBS. Detection antibody: 83972-1-PBS. Standard: Eg1118. Range: 0.313- 40 ng/mL.

Biolayer interferometry (BLl) kinetic assay of 83972-1-PBS against Mouse CD40/TNFRSF5 was performed. The affinity constant is 63.2 nM.