Methodology:
All living cells contain ATP and when the cell dies ATP is degraded by intracellular enzymes to ADP and finally AMP. The intracellular ATP concentration is similar in different cell types. This means that bacterial cells contain 1-2 attomoles per cell while animal and plant cells contain 100 to 10 000 times as much ATP. Extracellular ATP may be degraded by adding ATP consuming enzymes or by physical separation. Determination of intracellular ATP can then be used to estimate biomass. If the amount of ATP per cell is known the cell number may also be estimated. In sample which like urine may contain, extracellular ATP, ATP in somatic cells and ATP in bacterial cells all three ATP pools can be determined. Determination of bacterial ATP in urine is used to determine bacteriuria in the diagnosis of urinary tract infection. All ATP assays shall be calibrated by measuring the light before and after adding a known amount of ATP standard at the end of each test. This calibration obviates analytical interference from all sorts of variation of reagents and sample matrices and provides a result expressed in moles rather relative light units.

Intended use:

  1. Determination of bacterial ATP in urine in the diagnosis of urinary tract infection (UTI)1, 25, 35, 46, 47, 54.
  2. Antibiotic susceptibility testing5
  3. Antibiotic concentration determination
  4. Determination of somatic cells in cow milk (mastitis testing)
  5. Tumor chemosensitivity testing59
  6. Cell proliferation and cytotoxicity31, 36

Suitable products for bacteria: ATP Biomass Kit HS, Intracellular ATP Kit HS and Microbial ATP Kit HS

Suitable products for other cells: Cellular ATP Kit HTS, Cell Viability Kit SL

Recommended papers:

Thore A, Nilsson L, Hojer H, Ansehn S, Brote L. Effect of ampicillin on intracellular levels of adenosine triphosphate in bacterial cultures related to antibiotic susceptibility. Acta Pathol Microbiol Scand Sect B. 1977;85: 161–166.

Nilsson L (1978) New rapid bioassay of gentamicin based on luciferase assay of extracellular ATP in bacterial cultures. Antimicrob Agents Chemother 14:812–816

Nilsson L E, Hoffner S E, Ansehn S. Rapid susceptibility testing of Mycobacterium tuberculosis by bioluminescence assay of mycobacterial ATP. Antimicrob Agents Chemother. 1988;32:1208–1212.

Lundin, A., Hasenson, M., Persson, J. and Pousette, Å. (1986) Estimation of biomass in growing cell lines by adenosine triphosphate assay. Methods in Enzymology 133, 27-42.

Hasenson, M., Hartley-Asp, B., Kihlfors, C., Lundin, A. Gustafsson, J. -Å. and Pousette, Å. (1985) Effects of hormones on growth and ATP content of a human prostatic carcinoma cell line, LNCaP-r. The Prostate 7, 183-194.

Please choose the kit best describing your assay.

155-050

Cellular ATP

3968kr

For analyzing  mammalian cell samples as HTS

Cellular ATP Kit HTS is intended for use with mammalian cell samples. It is an easy-to-use kit where the lysing buffer, an ATPase inhibitor, and the ATP Reagent are combined in one reagent. The light emission is extremly stable, providing that all ATPases in the sample have been completly inhibited. That makes the 155-050 Cellular ATP Kit HTS suitable for high throughput screening.

In samples with a high density of ATPases, e.g. nerve cells, the ATPase inhibitor in the Cellular ATP Kit might not completly inhibit the ATPases. This will result in a slight decay of light. In such cases, a luminometer with dispensers could be used. Alternatively use 188-441 Cell Viability Kit SL which contains a stronger ATPase inhibitor.

 

188-441

Cell viability Kit SL

6943kr

For analyzing cell samples with high cell density

Cell Viability Kit SL is intended for use with mammalian cell samples with high cell density or with cell samples with higher density of ATPases, e.g. nerve cells. The kit contains a stronger ATPase inhibitor than 155-050 Cellular ATP Kit HTS and produces stable light.

A mammalian cell contains approx. 1 femtomole ATP. 188-441 Cell Viablity Kit SL is useful for quantification of ATP in the range 10^-11– 10^-7 mole/L. This kit can be used for determination of ATP levels in e.g. animal or bacterial cells in a variety of circumstances:
Estimation of cell numbers
Cell proliferation studies
Cytotoxicity studies

266-111

Intracellular ATP Kit HS

4125kr

Determination of intracellular ATP in bacterial cells

The sensitivity of this kit allows determination of low ATP levels in a variety of circumstances provided it is not important to degrade ATP in somatic cells:
1. Bacteriological control of liquids (e. g. drinking water, process water, beverages and beer)
2. Pharmaceuticals and cosmetics
3. Cell adhesion to surfaces
4. Quality assurance of laundry
5. Bacterial growth.
6. Antibiotic effects on bacteria

266-112

Microbial ATP Kit HS

4218kr

For determination of intracellular ATP in bacterial cells among mammalian cells

The sensitivity of this kit allows determination of low ATP levels in a variety of circumstances:
1. Bacteriological control of liquids (e. g. drinking water, process water, beverages and beer)
2. Pharmaceuticals and cosmetics
3. Cell adhesion to surfaces
4. Quality assurance of laundry
5. Bacterial growth.
6. Antibiotic effects on bacteria

266-311

ATP Biomass Kit HS

3365kr

For determination of the total ( both extracellular and intracellular) amount of ATP in a sample.

The ATP Biomass Kit is suitable for virtually all types of samples.

The sensitivity of this kit allows determination of low ATP levels in a variety of circumstances:
1 Liquids (e. g. drinking water, process water, environmental samples, beverages and beer)
2 Pharmaceuticals and cosmetics
3 Cell adhesion to surfaces (e. g. biocompatibility studies)
4 Quality assurance of laundry
5 Bacterial growth.
6 Antibiotic effects on bacteria