Performing Tests - QuenchGone21 Industrial

1. Total ATP (tATP)

The Total ATP test detects all ATP contained in the sample, inside and outside the cells. This is achieved by lysing the living cells and measuring their ATP as well as ATP that has already been released from dead cells.
TIP: High solids & complex fluids such as paints, coatings, adhesives, and slurries should be pre-diluted prior to analysis using the QG21I method. This ensures interference-free measurement of ATP. The recommended dilution is 1mL sample in 9mL sterile water or buffer. Use the same diluted sample to test both tATP and dATP when using the QG21I Standard (QG21I-50) test kit.

tATP Analyses consist of three steps:

Using a new wide-bore pipette tip, add 1mL of well-mixed sample to a new QG21I Extraction Tube. Cap and invert twice to mix. Allow at least 1 minute for incubation.

NOTE: At this point, the contents of the QG21I Extraction Tube can be capped and stored refrigerated between 2-8oC for up to 1 week prior to 2.2.
TIP: Wide-bore pipette tips are those that have sufficiently large openings to prevent tip plugging by sample particles. In general, standard 1-5mL and wide-mouth 100-1000µL pipette tips are sufficient for most samples. If required, increase the bore size of a tip using a clean pair of scissors.

Pour the QG21I Extraction Tube contents into a new QG21I Dilution Tube. Transfer the mixture back and forth between the two tubes at least three times for best mixing accuracy. Cap and invert twice to mix. Allow beads to settle


NOTE: At this point, the contents of the QG21I Dilution Tube are stable at room temperature for up to 4 hours.
TIP: If beads do not settle or settle slowly, tap the QG21I Dilution Tube gently to assist settling.
TIP: If the QG21I Extraction Tube cannot be poured into the QG21I Dilution Tube, simply pour the QG21I Dilution Tube contents into the QG21I Extraction Tube to liquefy its contents.

Using a new pipette tip, add 100µL of the QG21I Dilution Tube contents to a new 12x55mm test tube (the Assay Tube), add 2 drops (100µL) of Luminase, swirl gently five times, and insert into the Luminometer. Record RLUtATP for use in Step 4

NOTE: If RLUtATP < 10 on a Kikkoman Lumitester C-100 or C-110, you are below the low–detection limit. Report tATP (pg ATP/mL) = 0 in Step 4.
TIP: If “Scale Over” is returned, repeat the tATP analysis using 100µL of sample in 2.1 (EXTRACTION) and modify the dilution factor in Step 4 as noted.

2. Dissolved ATP (dATP)

The Dissolved ATP test uses the specialized LumiSolve dilution buffer to stabilize extracellular ATP (from dead and stressed cells) and also makes bound, or “complexed” ATP soluble. This allows ATP from outside of living cells to be measured without harming living cells.
TIP: Follow the same guidelines for sample pre-dilution as those described in Step 2.

dATP Analyses consist of two steps:

Using a new wide-bore pipette tip, add 1mL of well-mixed sample to a QG21I LumiSolve Tube. Cap and invert twice to mix. Allow at least 1 minute for incubation.

NOTE: The QG21I Dilution Tube can be capped and stored between 2 to 8oC for up to 1 week prior to 3.2.
TIP: Wide-bore pipette tips are those that have sufficiently large openings to prevent tip plugging by sample particles. In general, standard 1-5mL and wide-mouth 100-1000µL pipette tips are sufficient for most samples. If required, increase the bore size of a tip using a clean pair of scissors.

Using a new pipette tip, add 100µL of the QG21I LumiSolve Tube contents to a new 12x55mm test tube (the Assay Tube), add 2 drops (100µL) of Luminase, swirl gently five times, and insert into the Luminometer. Record RLUdATP for use in Step 4.

NOTE: If RLUdATP < 10 on a Kikkoman Lumitester C-100 or C-110, you are below the low–detection limit. Report dATP (pg ATP/mL) = 0 in Step 4.
TIP: If “Scale Over” is returned, repeat the dATP analysis using 100µL of sample in 3.1 (DILUTION) and modify the dilution factor in Step 4 as noted.