Description: HIV-1 p24 Capture ELISA.
Storage: Store Components as specified at 4oC.
Stability: Please see kit label.
Note: Read instructions and familiarize yourself with kit components before performing assays with this kit.
A) Plate holder with solid phase ligand strips (2 x 96 well, Store at 4oC)
B) Wash Buffer 10 X, (dilute 1:10 with deion-ized water)
C) Diluention Buffer (Store at 4oC)
D) p24 Standard (10ug/ml - 0.1ml, dilute before use, Store at 4oC)
E) Detector Reagent (dilute before use, Store at 4oC)
F) Substrate (Store at 4oC)
G) Stop Solution (Store at 4oC)
Bring Diluent Buffer (Component C) to room temperature before use.
1. Prepare several dilutions of p24 positive references (Component D) in diluent buffer (Component C).
Concentration range: 5ng/ml to 0.1ng/ml, 0.1ml/well. Concentration range 10 to 500 pg.
2. Prepare test samples using diluent buffer (Component C) in 50ng/ml range. Unknown samples (serum) may be used in 1:2 to 1:10 dilution range - use Component C .
3. Remove plate holder with strip from the cover. Align strip holder on the grid so that the markings read A-H Down, 1-12 Right.
4. Pipette 100µl of each positive reference (Step 1 above) and test samples (Step 2 above) into wells. Note position on the grid. Tap holder gently against table top to dislodge any air bubbles in wells. Cover plate with adhesive film and leave at room temperature for 1 hour. Note position of positive references and unknown samples on the grid diagram below.
5. Dump contents of wells and wash 3 times with 1 x wash buffer (Component B: 1:10 diluted with De-ionized H2O). Leave last wash in wells.
1. Dilute detector reagent (Component E) 1:100 in diluent buffer (Component C). Dump contents of wells (step 5 above) and pipette 100µl of diluted detector reagent (Component E) into each well. Cover plate and leave at room temperature for 1 hour.
2. Dump contents of wells, and wash wells 3 times with 1 x wash buffer (Component B). Dump last wash before continuing to the next step.
3. Add 100µl TMB substrate (Component F) to each well.