Annexin V Apoptosis Kit [FITC]

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Flow Cytometry: Annexin V Apoptosis Kit [FITC] [NBP2-29373] - Controls run before the experimental samples are analyzed, to adjust the fluorescent compensation on the flow cytometer due to the FITC signal from the ...read more
Flow Cytometry: Annexin V Apoptosis Kit [FITC] [NBP2-29373] - Adherent RAW cells treated with actinomycin-D, detached with a cell detachment solution and stained with the Annexin V Apoptosis Kit [FITC].

Product Details

Summary
Reactivity Hu, Mu, RtSpecies Glossary
Applications Flow, Flow-CS, ICC/IF
Conjugate
FITC

Order Details

Annexin V Apoptosis Kit [FITC] Summary

Description
Annexin V Apoptosis Kit [FITC] can identify apoptosis at an earlier stage than kits based on DNA fragmentation in the nucleus. However, it is like most assays and has limitations. Since Annexin V staining precedes the loss of membrane integrity which accompanies the later stages of cell death resulting from either apoptotic or necrotic processes, staining with Annexin V-FITC is typically used in conjunction with a live/dead dye such as propidium iodide (PI) or 7-Amino-Actinomycin (7-AAD) to allow the investigator to identify early apoptotic cells (PI negative, Annexin V-FITC positive) from dead cells (PI positive, AnnexinV-FITC positive). Viable cells with intact membranes exclude PI, whereas the membranes of dead and damaged cells are permeable to PI. For this reason, Annexin V staining has to be performed on live cells as opposed to assays which require para- formaldehyde/ethanol fixed cells.

The Annexin V assay works in the following manner: Cells that are viable are both Annexin V-FITC and PI negative. While cells that are in early apoptosis are Annexin V-FITC positive and PI negative and cells that are in late apoptosis or already dead are both FITC Annexin V and PI positive. This assay does not distinguish between cells that have undergone apoptotic death versus those that have died as a result of a necrotic pathway because the dead cells will stain with both Annexin V-FITC and PI (see image). However, when apoptosis is measured over time, cells can be often tracked from Annexin V- FITC and PI negative (viable, or no measurable apoptosis), to Annexin V-FITC positive and PI negative (early apoptosis, membrane integrity is present) and finally to Annexin V- FITC and PI positive (end stage apoptosis and death). The movement of cells through these three stages suggests apoptosis. In contrast, a single observation indicating that cells are both Annexin V-FITC and PI positive, in of itself, reveals less information about the process by which the cells underwent their demise. For this reason, it is a good idea to analyze samples from multiple time points.
Kit Type
Apoptosis Kit
Gene
ANXA5

Applications/Dilutions

Application Notes
Use in Flow cell surface reported in scientific literature (PMID 23955790) Use the Control Cells provided to set up compensation and quadrants. The Control Cells are positive for both Annexin V-FITC and PI.
The basal level of apoptosis and necrosis varies considerably within a give cell population. Even in the absence of induced apoptosis, most cell populations will contain a minor percentage of cells that are positive for apoptosis ((Annexin V-FITC positive, PI negative) and dead, necrotic, or in the late stages of apoptosis (Annexin V-FITC positive, PI positive). Thus an untreated cell population is used to define the basal level of apoptotic and dead cells.
Determine the percentage of cells that have been induced to undergo apoptosis by subtracting the percentage of apoptotic cells in the untreated from the treated population. Since cell death is the eventual outcome of cells undergoing apoptosis, cells in the late stages of apoptosis will have a damaged membrane and stain positive for PI as well as for Annexin V-FITC.
Consequently cells which have undergone necrosis are not distinguishable from those which have undergone apoptosis. Use in Immunocytochemistry/immunofluorescence reported in scientific literature (PMID 25947082). Use in FLOW cytometry reported in scientific literature ( PMID 27448441)
Publications
Read Publications using
NBP2-29373 in the following applications:

Reactivity Notes

Human reactivity reported in scientific literature (PMID: 23955790) Mouse reactivity reported in scientific literature (PMID: 22566271). Rat reactivity reported in scientific literature (PMID: 25947082)

Packaging, Storage & Formulations

Storage
Store at 4C. Do not freeze.

Kit Components

Components
  1. Compensation Control Cells
  2. Annexin V-FITC
  3. 10X PBS Solution
  4. 10X Binding Buffer
  5. Propidium Iodide (PI) Solution

Notes

ADDITIONAL ITEMS REQUIRED (NOT INCLUDED IN THE KIT)
Distilled H2O
Flow Cytometer
Experimental Cells
Centrifuge
Positive Control Cell Line (recommended, not required)

Alternate Names for Annexin V Apoptosis Kit [FITC]

  • Anchorin CII
  • annexin A5
  • Annexin V
  • annexin-5
  • ANX5
  • ANXA5
  • ANXV
  • Calphobindin I
  • Endonexin II
  • ENX2CBP-I
  • Lipocortin V
  • PAP-I
  • Placental anticoagulant protein 4
  • Placental anticoagulant protein I
  • PP4
  • Thromboplastin inhibitor
  • VAC-alpha
  • Vascular anticoagulant-alpha

Background

Apoptosis is the term that describes programmed cell death. It is believed to take place in the majority of animal and plant cells. It is a distinct event that triggers characteristic morphological and biological changes in the cellular life cycle. One of the early events that happens in the apoptotic pathway is the loss of plasma membrane asymmetry. In apoptotic cells, the membrane phospholipid phosphatidylserine (PS) is translocated from the inner to the outer leaflet of the plasma membrane, thereby exposing PS to the external cellular environment. Annexin V when labeled with a fluorescent tag, such as FITC, can be used with flow cytometry to measure this event. Annexin V is a 35-36 kDa Ca2+ dependent phospholipid-binding protein that has a high affinity for PS, and binds to cells with exposed PS.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Kits are guaranteed for 6 months from date of receipt.

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Publications for Annexin V Kit (NBP2-29373)(20)

We have publications tested in 3 confirmed species: Human, Mouse, Rat.

We have publications tested in 5 applications: FLOW, Flow, Flow - CS, Flow-CS, ICC/IF.


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FLOW
(8)
Flow
(1)
Flow - CS
(1)
Flow-CS
(5)
ICC/IF
(2)
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Human
(14)
Mouse
(2)
Rat
(2)
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Showing Publications 1 - 10 of 20. Show All 20 Publications.
Publications using NBP2-29373 Applications Species
Hartung F, Pardo LA. Guiding TRAIL to cancer cells through Kv10.1 potassium channel overcomes resistance to doxorubicin Eur. Biophys. J. 2016 Jun 27 [PMID: 27350552] (ICC/IF, Human) ICC/IF Human
Narendrula R. rRNA Disruption: A Predictive Marker of Response to Taxane Chemotherapy. Thesis 2014 (Flow-CS, Human)

Details:
A2789 human ovarian carcinoma cell line, Fig 12.
Flow-CS Human
Song W, Wang F, Lotfi P et al. 2-Hydroxypropyl-B-Cyclodextrin Promotes TFEB-mediated Activation of Autophagy: Implications for Therapy. J. Biol. Chem. 2014 Feb 20 [PMID: 24558044] (Flow-CS, Human) Flow-CS Human
Kumar A, Kant S, Singh SM. Targeting monocarboxylate transporter by alpha-cyano-4-hydroxycinnamate modulates apoptosis and cisplatin resistance of Colo205 cells: implication of altered cell survival regulation. Apoptosis 2013 Dec [PMID: 23955790] (Flow-CS, Human) Flow-CS Human
Vishvakarma NK, Kumar A, Singh V et al. Hyperglycemia of tumor microenvironment modulates stage-dependent tumor progression and multidrug resistance: implication of cell survival regulatory molecules and altered glucose transport. Mol Carcinog 2013 Dec [PMID: 22566271] (Flow-CS, Mouse) Flow-CS Mouse
Saydam O, Senol O, Wurdinger T et al. miRNA-7 attenuation in Schwannoma tumors stimulates growth by upregulating three oncogenic signaling pathways. Cancer Res. 2011 Feb 1 [PMID: 21156648] (Flow - CS, Human)

Details:
Flow (cell surface): Human primary Schwannoma and HEI-193 cell lines, S1.
Flow - CS Human
Badr CE, Niers JM, Morse D et al. Suicidal gene therapy in an NF-kB-controlled tumor environment as monitored by a secreted blood reporter. Gene Ther. 2011 May [PMID: 21150937] (Human)

Details:
Human glioma (Gli36) cells infected with lenti-NF-CU-IGluc and treated with TNF-alpha, 5-FC, or 5FC+TNF alpha, Fig 2D.
Human
Lu L, Wang S, Zheng L et al. Amyotrophic lateral sclerosis-linked mutant SOD1 sequesters Hu antigen R (HuR) and TIA-1-related protein (TIAR): implications for impaired post-transcriptional regulation of vascular endothelial growth factor. J Biol Chem. 2009 Dec 4 [PMID: 19805546]
Khurana S, Hollingsworth A, Piche M. Antiapoptotic Actions of Methyl Gallate on Neonatal Rat Cardiac Myocytes Exposed to H2O2. Oxidative Medicine and Cellular Longevity. 2014 Jan 12 (Rat)

Details:
Neonatal rat cardiac myocytes, Fig 3.
Rat
Skivka LM, Fedorchuk OG, Bezdeneznykh NO. The effect of antineoplastic drug NSC631570 on immunogenicity of B16 melanoma. J Exp Integr Med 2014 Mar 24 (Flow-CS, Mouse)

Details:
Metastatic mlanoma cells, Fig 2.
Flow-CS Mouse
Show All 20 Publications.

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Bioinformatics

Gene Symbol ANXA5
Entrez