Annexin V-APC/DAPI Apoptosis Kit

Catalog Number: abs50004

Price:

$285.00 USD

Size: 50T 100T

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Product Details

Product Specification

Usage

Experimental procedure:
1. Annexin V-APC and cell incubation
1. Induce apoptosis according to the experimental protocol. Centrifuge at 300 g for 5 min, discard supernatant, collect cells, gently resuspend cells with PBS and count.
2. Centrifugation collection 1-5 × 10⁵The supernatant was discarded. The cells were washed once with PBS and the supernatant was discarded after centrifugation.
3. Resuspend the cells with 500 µ L of 1X Binding Buffer.
4. Add 5 µ L Annexin V-APC and 5 µ L DAPI.
5. Gently vortex and mix well, and incubate at room temperature in the dark for 15 minutes.
6. Test immediately. If it cannot be detected in time, please put it on ice to protect from light and complete the test within 1 hour.
2. Flow cytometry quantification
Flow cytometry checks the available APC channel for Annexin V-APC, and the DAPI channel is preferred for DAPI detection, followed by the PacificBlue channel. For adherent cells, the cell culture medium was first collected, the cells were washed once with PBS, and the cells were digested with trypsin, then the collected cell culture medium was added, the cells were gently blown down, the supernatant was discarded by centrifugation, and the cells were resuspended with PBS and counted, and then step 1 2-5 was performed.
3. Fluorescence microscope detection The cell suspension in step 1.5 is coated on a glass slide. Cover cells with a coverslip. If adherent cells are analyzed, the cells are cultured directly on the cell slide. After incubation (step 1.5), the cell slide was inverted on a glass slide and the cells were observed.

Beilstein

Description

Annexin V-APC/DAPI Apoptosis Detection Kit can detect the apoptosis of suspended cells and adherent cells.
Annexin V is a calcium-dependent phosphatidylserine binding protein with high affinity for phosphatidylserine PS, and Annexin V-APC can bind to the cell membrane of early apoptotic cells through PS exposed extracellularly. Apoptosis can be detected by flow cytometry or fluorescence microscopy.
Due to late apoptosis or loss of membrane integrity in necrotic cells, 4 ', 6-Diamidino-2-phenylindole dihydrochloride (4', 6-Diamidino-2-Phenylindole, DAPI) can specifically bind to double-stranded DNA and produce strong fluorescence. When used in conjunction with AnnexinV, it can distinguish cells in different apoptotic stages.
Product composition:

Name	20T	50T	100T
Annexin V-APC	100 μL	250 μL	500 μL
1X Binding Buffer	10mL	25mL	50mL
DAPI	100 μL	250 μL	500 μL

PubChem CID

General Notes

1. It should be tested as soon as possible after dyeing. Too long time may lead to increased number of apoptotic or necrotic cells.
2. When detecting adherent cells, the suspension cells produced after apoptosis induction should be collected and detected together with the adherent cells collected subsequently.
3. Try to avoid mechanical damage caused by digestive adherent cells. At the same time, the digestive juice of pancreatic enzymes should be as free of EDTA as possible, as EDTA affects the binding of Annexin V to phosphatidylserine.
4. If pancreatic enzyme containing EDTA is used, the cells should be thoroughly cleaned after collection to ensure that EDTA is removed.
5. Fluorescent substances are easy to quench. When performing fluorescence observation, try to shorten the observation time, and try to pay attention to light preservation during operation and storage.
6. For your safety and health, please wear a lab coat and disposable gloves.

Storage Temp.

Store at 2-8 °C, do not freeze, shelf life 1 year.

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Annexin V-APC/DAPI Apoptosis Kit