Methods in Anopheles Research - MR4

Chapter 8 : Field Techniques8.2 Plasmodium falciparum Sporozoite ELISAPage 5 of 12Dilutions for Positive Control Antigens for CS-ELISASerial dilutions are easily prepared by one of the two following methods:1. All working concentrations required can be prepared and labeled ahead of time. When addition ofpositive controls to wells of ELISA plate is required, 50l of each solution is added. This method issuited for inexperienced pipetters.Label microfuge tubes with solution number and prepare sequentially according to chart. Remainingvolumes can be stored at -20ºC and used again when necessary.Solution Volume of Positive Control Volume of BlockingNumber AntigenBuffer (l)PlasmodiumspeciesAntigenConcentration(pg/50l)P. falciparum Stock Lyophilized Pf 1000 500,000I 20l of stock 1000 5,000*II 10l of I 500 100*III 500l of II 500 50*IV 500l of III 500 25*V 500l of IV 500 12*VI 500l of V 500 6*VII 500l of VI 500 3*VIII 500l of VII 500 1.5*IX 0 (Blank) 500 0 (Blank)P. vivax 210 Stock Lyophilized Pv210 500 500,000A 10l of stock 1000 5,000B 20l of A 500 200*C 200l of B 800 40*D 500l of C 500 20*E 500l of D 500 10*F 500l of E 500 5*G 500l of F 500 2.5*H 500l of G 500 1.25* I 500l of H 500 0.6*J 0 (Blank) 500 0P. vivax 247 Stock Lyophilized Pf 1000 227,500*A 20l of stock 1000 4,450*B 500l of I 500 2,275*C 500l of II 500 1,140*D 500l of III 500 570*E 500l of IV 500 285*F 500l of V 500 140*G 500l of VI 500 70*H 0 (Blank) 500 0 (Blank)* Amounts used in ELISA plate wells for the quantitative assays