Annexin V-Cy3 Apoptosis Kit Plus

Details zu Produkt Nr. ABIN412001
Antigen
  • anx
  • anx5
  • ANX V
  • anxa5
  • cb989
  • wu:fa98f06
  • wu:fj10f10
  • MGC89158
  • ANX5
  • ENX2
  • PP4
  • RPRGL3
  • Anx5
  • R74653
  • LC5
  • enx2
  • annexin A5
  • annexin A5b
  • Annexin A5
  • annexin A5 L homeolog
  • ANXA5
  • anxa5b
  • anxa5
  • Anxa5
  • anxa5.L
Reaktivität
Chemical
12
10
7
6
6
5
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3
2
2
2
1
1
1
1
Nachweismethode
Fluorometric
Applikation
Apoptosis Detection (AD), Fluorescence Microscopy (FM), Flow Cytometry (FACS)
Optionen
Verwendungszweck The assay is based on the observation that soon after initiating apoptosis, cells translocate the membrane phospholipid phosphatidylserine (PS) from the inner face of the plasma membrane to the cell surface. Once on the cell surface, PS can easily be detected by staining with a fluorescent conjugate of Annexin V, a protein that has a strong natural affinity for PS. The one-step staining procedure takes only 10 minutes. In addition, the assay can be directly performed on live cells, without the need of fixation. The Annexin V-Cy3 Apoptosis Detection Kit Plus includes annexin V-Cy3, SYTOX green dye, and binding buffer. The SYTOX green dye is impermeant to live cells and apoptotic cells, but stains necrotic cells with intense green fluorescence by binding to cellular nucleic acids. After staining a cell population with annexin V-Cy3 and SYTOX Green dye in the provided binding buffer, apoptotic cells show red fluorescence, dead cells show green fluorescence and live cells show little or no fluorescence. These populations can easily be distinguished by Fluorescence microscopy using FITC and rhodamine filters or by flow cytometry using the FL1 channel (Ex. 488 nm/Em. 530 nm) for SYTOX Green dye and FL2 channel for Annexin V-Cy3 (Ex. 543 nm/Em. 570 nm).
Proben Cell Samples
Nachweismethode Fluorometric
Spezifität The assay is based on the observation that soon after initiating apoptosis, most cell types translocate the membrane phospholipid phosphatidylserine (PS) from the inner face of the plasma membrane to the cell surface. Once on the cell surface, PS can easily be detected by staining with a fluorescent conjugate of Annexin V, a protein that has a strong natural affinity for PS.
Produktmerkmale Annexin V-Cy3 Apoptosis Detection Kit Plus: Simple One Step Assay to Detect Apoptotic, Necrotic & live cells within 10 min. No need to fix the cells.
Detection: Flow Cytometry or fluorescence microscopy
The one-step staining procedure takes only 10 minutes. In addition, the assay can be directly performed on live cells, without the need of fixation. The Annexin V-Cy3 Apoptosis Detection Kit Plus includes annexin V-Cy3, SYTOX green dye, and binding buffer. The SYTOX green dye is impermeant to live cells and apoptotic cells, but stains necrotic cells with intense green fluorescence by binding to cellular nucleic acids. After staining a cell population with annexin V-Cy3 and SYTOX Green dye in the provided binding buffer, apoptotic cells show red fluorescence, dead cells show green fluorescence and live cells show little or no fluorescence. These populations can easily be distinguished by Fluorescence microscopy using FITC and rhodamine filters or by flow cytometry using the FL1 channel (Ex. 488 nm/Em. 530 nm) for SYTOX Green dye and FL2 channel for Annexin V-Cy3 (Ex. 543 nm/Em. 570 nm).
Bestandteile Annexin V-Cy3
SYTOX Green Dye
Binding Buffer
Protein- /Substanz
Andere Bezeichnung Annexin V (ANXA5 ELISA Kit Abstract)
Substanzklasse Chemical
Hintergrund The assay is based on the observation that soon after initiating apoptosis, cells translocate the membrane phospholipid phosphatidylserine (PS) from the inner face of the plasma membrane to the cell surface. Once on the cell surface, PS can easily be detected by staining with a fluorescent conjugate of Annexin V, a protein that has a strong natural affinity for PS. The one-step staining procedure takes only 10 minutes. In addition, the assay can be directly performed on live cells, without the need of fixation. The Annexin V-Cy3 Apoptosis Detection Kit Plus includes annexin V-Cy3, SYTOX green dye, and binding buffer. The SYTOX green dye is impermeant to live cells and apoptotic cells, but stains necrotic cells with intense green fluorescence by binding to cellular nucleic acids. After staining a cell population with annexin V-Cy3 and SYTOX Green dye in the provided binding buffer, apoptotic cells show red fluorescence, dead cells show green fluorescence and live cells show little or no fluorescence. These populations can easily be distinguished by Fluorescence microscopy using FITC and rhodamine filters or by flow cytometry using the FL1 channel (Ex. 488 nm/Em. 530 nm) for SYTOX Green dye and FL2 channel for Annexin V-Cy3 (Ex. 543 nm/Em. 570 nm).
Applikations-hinweise Detecting apoptosis in living cells by flow cytometry or fluorescence microscopy. Cy3 shows brighter red fluorescence.
Kommentare

Further details regarding sample type: Live cells

Testdauer 0.5 h
Protokoll 1. Induce apoptosis by desired method. Concurrently incubate a control culture without induction.
2. Collect 1-5 x 10^5 cells by centrifugation.
3. Resuspend cells in 500 µL of 1X Binding Buffer.
4. Add 5 µL of Annexin V-Cy3 and 1 µL of SYTOX Green dye.
Note: Thaw the SYTOX Green dye in room temperature before use.
5. Incubate at room temperature for 5-10 min in the dark.
6. Analyze the stained cells by flow cytometry using FL1 channel for SYTOX Green dye (Ex = 488 nm, Em = 530 nm) and FL2 channel for Annexin V-Cy3 (Ex = 543 nm, Em = 570 nm).
The cell population should separate into three groups: live cells with only a low level of fluorescence, apoptotic cells with red fluorescence and necrotic cells with green fluorescence.
The flow cytometric results can also be confirmed by viewing the cells under a fluorescence microscope using FITC filter for SYTOX and rhodamine filter for Annexin V-Cy3.
For adherent cells, gently trypsinize and wash cells once with serum-containing media before incubation with Annexin V-Cy3 and SYTOX dye.
Beschränkungen Nur für Forschungszwecke einsetzbar
Handhabung Protect from light. Product is photosensitive.
Lagerung 4 °C
Haltbarkeit 12 months
Produkt verwendet in: Morrow, Grant, Mujawar, Dubrovsky, Pushkarsky, Kiselyeva, Jennelle, Mukhamedova, Remaley, Kashanchi, Sviridov, Bukrinsky: "Stimulation of the liver X receptor pathway inhibits HIV-1 replication via induction of ATP-binding cassette transporter A1." in: Molecular pharmacology, Vol. 78, Issue 2, pp. 215-25, 2010 (PubMed).

Boehm, Sen, Seethala, Gooding, Freilino, Wong, Wang, Johnson, Grandis: "Combined targeting of epidermal growth factor receptor, signal transducer and activator of transcription-3, and Bcl-X(L) enhances antitumor effects in squamous cell carcinoma of the head and neck." in: Molecular pharmacology, Vol. 73, Issue 6, pp. 1632-42, 2008 (PubMed).

Lin, Lee, Hung, Li, Yang-Yen, Yen: "Survival factor withdrawal-induced apoptosis of TF-1 cells involves a TRB2-Mcl-1 axis-dependent pathway." in: The Journal of biological chemistry, Vol. 282, Issue 30, pp. 21962-72, 2007 (PubMed).

Sok, Coppelli, Thomas, Lango, Xi, Hunt, Freilino, Graner, Wikstrand, Bigner, Gooding, Furnari, Grandis: "Mutant epidermal growth factor receptor (EGFRvIII) contributes to head and neck cancer growth and resistance to EGFR targeting." in: Clinical cancer research : an official journal of the American Association for Cancer Research, Vol. 12, Issue 17, pp. 5064-73, 2006 (PubMed).

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