Application
This kit can be used to measure the ATPase activity in samples, such as cell membranes, mitochondria, microsomes samples.
Detection significance
ATPase exists on cell membrane and organelle membrane. It is a protease in biomembrane. It plays an important role in material transport, energy conversion and information transmission.
Detection principle
ATPase can decompose ATP to produce ADP and inorganic phosphorus. The activity of ATPase can be calculated indirectly by measuring the content of inorganic phosphorus.
Experimental instruments
Tube, Micropipettor, Vortex mixer, 37℃ thermostat water bath, Spectrophotometer (660 nm)
Operation steps
1. Enzymatic reaction
| Tube A | Tube B | Tube C | Tube D | Tube E |
Reagent 1 (μL) | 70 | 70 | 50 | 70 | 50 |
Reagent 2 (μL) | 20 | 20 | 20 |
| 20 |
Reagent 3 (μL) |
|
|
| 20 | 20 |
Reagent 4 (μL) | 20 | 20 | 20 | 20 | 20 |
Reagent 5 (μL) |
|
| 20 | 20 | 20 |
Reagent 6 (μL) | 20 | 20 | 20 |
|
|
Sample (μL) |
| 100 | 100 | 100 | 100 |
Mix fully and react accurately in 37℃ water bath for 10 min. | |||||
Reagent 7 (μL) | 50 | 50 | 50 | 50 | 50 |
Sample (μL) | 100 |
|
|
|
|
Mix fully and centrifuge at 3000~4000 rpm for 10 min, then take 100 μL supernatant for phosphorus determination. | |||||
2. Phosphorus determination
| Standard tube | Tube A | Tube B | Tube C | Tube D | Tube E |
1 μmol/mL Phosphorus standard application solution (μL) | 100 |
|
|
|
|
|
Supernatant (μL) |
| 100 | 100 | 100 | 100 | 100 |
Phosphorus determination solution (μL) | 2000 | 2000 | 2000 | 2000 | 2000 | 2000 |
Mix fully and incubate in 45℃ water bath for 20 min, then cool down to room temperature.Set the spectrophotometer to zero with double-distilled water and measure the OD value of each tube at 660 nm with 1 cm diameter cuvette. | ||||||
Note: 1. Tube A is contrast tube; Tube B is Na+/k+ -ATPase tube; Tube C is Mg2+-ATPase tube. Tube D is Ca2+-ATPase tube; Tube E is Ca2+/Mg2+-ATPase tube. (Please choose corresponding tube number to do experiment according to requirement.).
2. Reference sampling:
Erythrocyte membrane: 100 μL, 2% tissue cell membrane: 100 μL,
2% mitochondria: 100 μL, 2% micro mitochondrion: 100-200 μL
Notes
1. This kit is for research use only.
2. Please progress strictly with operation procedures.
3. Do not use components from different batches of kit.
4. The valid period of kit is 6 months and the expiration date is on the packing box.
상품문의가 없습니다.
추가비용을 고객에게 부담시지키 않습니다. (단, 고객 변심 또는 주문 반복으로 인한 경우의 반환비용은 고객님이 부담하셔야
합니다.)
::: 교환 및 반품이 가능한 경우:::
단, 상품을 개봉하여 상품가치가 상실된 경우에는 교환/반품이 불가능합니다.
:::교환 및 반품이 불가능한 경우:::
주문 취소 및 반품으로 환불을 요청하실 경우에는 E-mail(celgen-bio@celgen-bio.com)이나 고객만족센터 (042-824-9026)을
통해 요청하시면 친절하게 처리해 드리겠습니다.
주문 취소 후 반품 가능 여부를 확인한 다음 3일 이내에 결제 금액을 환불해 드리겠습니다.