One-step TUNEL In Situ Apoptosis Kit (Green, FITC) (E-CK-A320)
- +5
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 fluorescence microscope. |
| Detection Method | Fluorometric method |
| Sample Type | Paraffin section;frozen section;cell slide |
| Assay Time | 3 hours |
| Detection Instrument | Fluorescence Microscope |
| Dye Type | FITC |
| Ex/Em | 490/520 |
| Channel Set | FITC |
| Filter Set | FITC |
| Other Reagents Required | 4% Paraformaldehyde(E-IR-R113),0.2% Triton X-100(E-IR-R122),PBS(E-BC-R187),ddH2O,Mounting solution with anti-fluorescence quencher(E-IR-R119),Xylene,Ethanol |
| 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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1 Results
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Q1:What thickness do you recommend for frozen slices?
Generally, the thickness of frozen slices is 8-10 µm. If the slice is thick, the incubation time of protease K can be extended appropriately according to the actual situation.
Q2:What is the well state of dewaxing?
The tissue is transparent, and no water drop can be left on the slide.
Q3:What is the role of the TdT equilibration buffer in the TUNEL Kit?
The TdT equilibration buffer in the TUNEL kit is designed to provide optimal reaction conditions for the TdT enzyme.
Q4:What is the resolution ratio of CCK-8?
The best linearity of CCK-8 is between 0.2-2.5 (Optical Density).
Q5:What is the recommended amount of PBS washing?
PBS added during washing steps must completely cover the sample, and ensure the sample moist during the experiment to prevent the failure of the experiment caused by dry slides.
Q6: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.
Q7:What is the difference between TUNEL-fluorescence and HRP-DAB TUNEL?
Color development of DAB (result of TUNEL-HRP) is observed with an ordinary microscope, the results of TUNEL nuclear staining were served with white light. As for TUNEL fluorescence, the results of TUNEL and nuclear staining were observed with different fluorescence channels seperately with better definition and resolution.
Q8: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.Q9:What is the difference between one-step and two-step Annexin V operation?
Both systems have the same resolution ratio, while one-step operation takes easier process, and two-step operation requires less reagent.
Q10:What is the difference between FITC and Elab Fluor® 488 in TUNEL kit?
ElabFluor®488 offers better quenching resistance and is more suitable for longer microscopic observations than that of FITC.
