One-step TUNEL Flow Cytometry Apoptosis Kit (Green, FITC) (E-CK-A420)
For research use only. Order now, ship in 3 days
| Detection Principle | When cells undergo apoptosis, specific DNA endonucleases will be activated, cutting the genomic DNA between the nucleosomes. The DNA of apoptotic cells is cleaved into multimers of 180~200bp fragments, corresponding to the oligonucleosomal size. Therefore, the DNA of apoptotic cells typically migrates as a ladder of 180~200bp on an agarose gel. The exposed 3'-OH of the broken DNA can be catalyzed by Terminal Deoxynucleotidyl Transferase (TdT) with fluorescein labeled dUTP, which can be detected with flow cytometry. |
| Detection Method | Fluorometric method, Flow cytometry |
| Sample Type | Cell samples |
| Assay Time | 2.5-3 hours |
| Detection Instrument | Flow Cytometer |
| Dye Type | FITC |
| Ex/Em | 490/530 |
| Channel Set | FITC |
| Other Reagents Required | PBS buffer (with 1%BSA) (PH7.2-7.4), PBS (with 1% Saponin) (pH7.2-7.6), 4% Polyformaldehyde (dissolved in PBS)(E-IR-R113), Deionized water. |
| Storage | This product can be stored at -20°C for 12 months with shading light. |
| Expiration date | 12 months |
| Shipping | Ice bag |
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Q1:What is the resolution ratio of CCK-8?
The best linearity of CCK-8 is between 0.2-2.5 (Optical Density).
Q2:What is the purpose of using equilibrition buffer before labeling?
The equilibrition process is to make a better working condition of TdT enzyme that benefit a better staining result.
Q3:What is the difference between TUNEL in-situ kit and TUNEL flow cytometry kit?
The main difference lies in the detection methods, for TUNEL in-situ kits, a fluorescence microscope is used for observing the results, as for TUNEL flow ytometry kit, the sample will be detected by a flow cytometer.
On the other hand, TUNEL in-situ kit is more suitable for tissue sections and cell slides or smears, while TUNEL flow ytometry kit is more suitable for suspended cells and adherent cells.Q4:What are the reasons that lead to false positives in TUNEL?
Cells with high proliferative or metabolic active may have a break in the DNA strand, both cases may end up with false-positive result. Evaluation of cell morphology may be used if there are doubts about the interpretation of the results.
Q5:Is there any TUNEL assay kit that can detect apoptosis in erythrocyte?
The detection of TUNEL is based on DNA fragmentation in late stage apoptosis, so it is not avaliable for erythrocyte as there is no nuclei or other organelles containing DNA in erythrocyte.
Q6:Is the fluorochrome in Calcein AM/PI stable? How long can it be stored after staining?
Normally, it is better to detect in 1-2 hours after staining. If the cells required further incubation, it can be kept in CO2 incubator with no more than 48 hours. When using microscope, it is recommanded to turn the illumination down and extend exposure time.
Q7:Can TUNEL assay kit be used for mouse samples?
There is no species limination in TUNEL assay kits.
Q8:Can the One-step TUNEL kit detect a sperm sample?
Yes, it can be used to test sperm samples.
