• Enhanced ATP Assay Kit 200T
  • Enhanced ATP Assay Kit 200T

Enhanced ATP Assay Kit 200T

The Enhanced ATP Assay Kit can be used to detect ATP levels in ordinary solutions, cells, or tissues. Cell and tissue samples can be lysed in one step. Sample preparation, detection sensitivity up to 0.1nM, and chemiluminescence can remain stable for up to 30 minutes. The samples obtained from this kit can also be subjected to Western testing simultaneously.
$257.00
  • Enhanced ATP Assay Kit 200T

SPECIFICATION


                                          Enhanced ATP Assay Kit 200T

The Enhanced ATP Assay Kit can be used to detect ATP levels in ordinary solutions, cells, or tissues. Cell and tissue samples can be lysed in one step. Sample preparation, detection sensitivity up to 0.1nM, and chemiluminescence can remain stable for up to 30 minutes. The samples obtained from this kit can also be subjected to Western testing simultaneously.
ATP, as the most important energy molecule, plays an important role in various physiological and pathological processes of cells. Changes in ATP levels can affect cell function. Usually, ATP levels decrease when cells undergo apoptosis, death, or in some toxic state; High glucose stimulation can upregulate the intracellular ATP levels of certain cells. Usually, a decrease in ATP levels indicates impaired or decreased mitochondrial function, and a decrease in ATP levels during cell apoptosis usually occurs simultaneously with a decrease in mitochondrial membrane potential.
Technical principles:
This kit is developed based on the energy provided by ATP when firefly luciferase (also known as luciferase) catalyzes the production of fluorescence from luciferin. When both firefly luciferase and luciferin are in excess, the production of fluorescence is proportional to the concentration of ATP within a certain concentration range. This allows for highly sensitive detection of ATP concentration in the solution.
Product advantages:
1. Sample preparation is simple.
This kit provides ATP detection lysate that can be used for cell and tissue lysis. After simple lysis, it can be used for ATP detection. There is no need for tedious operations such as perchloric acid or trichloroacetic acid (TCA) extraction, or boiling after sample cracking.
2. High sensitivity, wide linear range, ranging from 0.1nM to 10 μ Good detection effect within the M range.
This reagent kit can detect ATP with concentrations as low as 0.1nM when the sample volume is 100 μ L. The concentration of ATP in conventional cell or tissue lysates is 0.1-1 μ M. The intracellular ATP level of some common cells is approximately 10 nmol/mg protein. And the detection concentration range of this reagent kit is very wide, with a detection limit of up to 10 μ M. And within 0.1nM-10 μ A good standard curve can be formed within the M range.
3. The reading is stable.
This reagent kit has been specially optimized to ensure very stable chemiluminescence during ATP detection. The detection results of the ATP standard curve showed that there was no significant decrease in chemiluminescence within 10 minutes after the start of the reaction, and the decrease in chemiluminescence within 30 minutes after the start of the reaction did not exceed 10%.
4. The prepared sample has good compatibility.
The cell or tissue samples obtained from the lysis of ATP detection lysate in this kit can not only be used for ATP detection, but also for protein concentration detection, SDS-PAGE, or some conventional Western detection of easily soluble proteins.
5. Convenient and fast to use.
Usually, 10-20 samples can be determined within 30-60 minutes.
Saving conditions:
-Stored at 20 ℃, valid for six months- Stored at 70 ℃, valid for one year. ATP detection reagents need to be stored away from light.
Instructions for use:
1. Preparation for sample determination: (Note: Sample cracking needs to be carried out at 4 ℃ or on ice)
For adherent cells:
Remove the culture medium by suction, and add 200 microliters of lysate to each well on a 6-well plate (equivalent to 1/10 of 2 milliliters of cell culture medium) to lyse the cells. In order to fully lyse cells, a pipette can be used to repeatedly blow or shake the culture plate to fully contact the lysate and lyse the cells. Usually, cells immediately lyse upon contact with the lysate. Centrifuge 12000 g at 4 ℃ for 5 minutes after cracking, and extract the supernatant for subsequent determination.
For suspended cells:
Centrifuge and precipitate cells using a centrifuge tube, discard the supernatant, gently scatter the cells, and add 200 microliters of lysate according to the amount of cells per well in a 6-well plate to lyse the cells. During cell lysis, in order to achieve sufficient lysis, the bottom of the centrifuge tube can be hit or appropriate Vortex can be used to fully contact the lysis solution and lyse the cell. Usually, cells immediately lyse upon contact with the lysate. Centrifuge 12000 g at 4 ℃ for 5 minutes after cracking, and extract the supernatant for subsequent determination.
For tissue samples:
Add approximately 100-200 microliters of lysate to every 20 milligrams of tissue, and then homogenize using a glass homogenizer or other homogenizing equipment. Adequate homogenization can ensure complete tissue lysis. Centrifuge 12000 g at 4 ℃ for 5 minutes after cracking, and extract the supernatant for subsequent determination.
2. Preparation for standard curve measurement:
Dissolve the reagent to be used on an ice bath and dilute the ATP standard solution with ATP detection lysate to an appropriate concentration gradient. The specific concentration needs to be determined based on the concentration of ATP in the sample. The initial test can detect 0.01, 0.03, 0.1, 0.3, 1, 3, and 10 μ In subsequent experiments, the concentration range of the standard substance can be adjusted appropriately based on the ATP concentration in the sample.
3. Preparation of ATP detection working solution:
Prepare an appropriate amount of ATP testing working solution according to the ratio of 100 microliters of ATP testing working solution required for each sample or standard. Dissolve the reagent to be used on an ice bath. Take an appropriate amount of ATP detection reagent and dilute it with ATP detection reagent diluent in a 1:4 ratio. For example, 1 milliliter of ATP detection reagent is added to 4 milliliters of ATP detection reagent diluent to prepare 5 milliliters of ATP detection working solution. The diluted ATP detection reagent is the ATP detection working solution used for subsequent experiments. ATP detection working solution can be temporarily stored on an ice bath.
4. Determination of ATP concentration:
a. Add 100 microliters of ATP detection working fluid to the detection hole or tube. Leave at room temperature for 3-5 minutes to consume all the background ATP, thereby reducing the background. You can add 100 microliters of ATP detection working fluid to each of the 10-20 detection wells or tubes at once, thereby saving time.
b. Add 20 microliters of sample to the testing hole or tube
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