Apoptosis assay cell type - HEK293

A gross majority of classical apoptotic attributes can be quantitatively examined by flow cytometry, the preferred platform for rapid assessment of multiple cellular attributes at a single-cell level. However, sample preparation for such flow cytometry-based techniques could be challenging. Cell harvesting by trypsinization, mechanical or enzymatic cell disaggregation from tissues, extensive centrifugation steps, may all lead to preferential loss of apoptotic cells. To overcome this strictly follow manufacturers instruction of the detection kit.

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Found 3 matching solutions for this experiment

ApopTag® Peroxidase In Situ Apoptosis Detection Kit

Millipore

Upstream tips	Protocol tips	Downstream tips
- Do not warm the stock reagents before dispensing them from containers. - Ethanol should not be substituted for n-butanol as it will destain nuclei.		- If staining of both nuclei and cytoplasm is apparent, the nonspecific component often can be mitigated by reducing the development time in substrate. - If a very high percentage of the nuclei in a tissue can be stained with TdT, but not without TdT, there appears to be staining of DNA only.

Upstream tips
- Do not warm the stock reagents before dispensing them from containers. - Ethanol should not be substituted for n-butanol as it will destain nuclei.
Downstream tips
- If staining of both nuclei and cytoplasm is apparent, the nonspecific component often can be mitigated by reducing the development time in substrate. - If a very high percentage of the nuclei in a tissue can be stained with TdT, but not without TdT, there appears to be staining of DNA only.

Manufacturer protocol Publication protocol 1 Relevant paper

FITC Annexin V Apoptosis Detection Kit I

BD Biosciences

Upstream tips	Protocol tips	Downstream tips
- 3×105 cells were seeded per dish. - FC101 (0–5 µM) is treated on cells for 72 h.	- Treated with FITC Annexin V and PI and incubated for 15 min at RT (25°C) in the dark

Upstream tips
- 3×105 cells were seeded per dish. - FC101 (0–5 µM) is treated on cells for 72 h.
Protocol tips
- Treated with FITC Annexin V and PI and incubated for 15 min at RT (25°C) in the dark

Manufacturer protocol Publication protocol 1 Relevant paper

Annexin V-FITC Apoptosis Detection Kit

Millipore

Upstream tips	Protocol tips	Downstream tips
- Cells were transfected and pre treated with CRH/Ucn2.	- After 24 h, the cells were used to assay apoptosis

Upstream tips
- Cells were transfected and pre treated with CRH/Ucn2.
Protocol tips
- After 24 h, the cells were used to assay apoptosis

Manufacturer protocol Publication protocol 1 Relevant paper

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