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)
Reviewed Applications
Read 2 Reviews rated 4
using
NBP2-29373 in the following application:

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

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)(27)

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

We have publications tested in 6 applications: CA, FLOW, Flow, Flow - CS, Flow-CS, ICC/IF.


Filter By Application
CA
(2)
FLOW
(8)
Flow
(2)
Flow - CS
(1)
Flow-CS
(5)
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(2)
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Human
(15)
Mouse
(3)
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(2)
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Showing Publications 1 - 10 of 27. Show All 27 Publications.
Publications using NBP2-29373 Applications Species
Husain S, Verma SK, Yasin D et al. Facile green bio-fabricated silver nanoparticles from Microchaete infer dose-dependent antioxidant and anti-proliferative activity to mediate cellular apoptosis Bioorganic chemistry Dec 11 2020 [PMID: 33341280]
Chiavari M, Ciotti GMP, Canonico F et al. PDIA3 Expression in Glioblastoma Modulates Macrophage/Microglia Pro-Tumor Activation Int J Mol Sci Nov 3 2020 [PMID: 33153019] (CA, Human)

Details:
Cell apoptosis analysis of CHME-5 cell line transfected with PDIA3 siRNA
CA Human
Xia W, Bai H, Deng Y, Yang Y PLA2G16 is a mutant p53/KLF5 transcriptional target and promotes glycolysis of pancreatic cancer J Cell Mol Med Sep 27 2020 [PMID: 32985124] (Flow, Mouse) Flow Mouse
Rybalchenko OV, Anisimova NY, Kiselevsky MV et al. The influence of ultrafine-grained structure on the mechanical properties and biocompatibility of austenitic stainless steels J. Biomed. Mater. Res. Part B Appl. Biomater. 2019 Oct 16 [PMID: 31617961]
Basset C, Bonnet-Magnaval F, Navarro MG, Touriol C et al. Api5 a new cofactor of estrogen receptor alpha involved in breast cancer outcome Oncotarget 2017 Aug 08 [PMID: 28881748] (Flow, Human) Flow Human
Chen Y, Huang H, Yao C et al. Antitumor activity of combined endostatin and thymidine kinase gene therapy in C6 glioma models Cancer Med 2016 Jul 01 [PMID: 27366865]
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
Huang CL, Chao CC, Lee YC et al. Paraquat Induces Cell Death Through Impairing Mitochondrial Membrane Permeability. Mol. Neurobiol. 2015 May 07 [PMID: 25947082] (ICC/IF, FLOW, Rat)

Details:
Annexin V-FITC-conjugated apoptosis kit was used for analyzing apoptotic cells in experiments involving PC12 cells. The cells with or without 24 hours of Paraquat/PQ treatments (+ dibucaine or cyclosporin A treatments) were subjected to Annexin V-FITC-staining followed by microscopgy imaging or flow cytometric analysis (Fig 1D).
ICC/IF, FLOW Rat
Xue Y, Wu L, Liu Y et al. ENTPD5 Induces Apoptosis in Lung Cancer Cells via Regulating Caspase 3 Expression PLoS ONE 2015 Mar 21 [PMID: 25794010] (FLOW, Human) FLOW Human
Song W, Soo Lee S, Savini M et al. Ceria nanoparticles stabilized by organic surface coatings activate the lysosome-autophagy system and enhance autophagic clearance. ACS Nano. 2014 Oct 28 [PMID: 25315655] (FLOW, Human)

Details:
Annexin V - FITC Apoptosis Kit (Imgenex's CytoGLO Annexin V-FITC Apoptosis Detection Kit 10085K) used for detecting - Annexin V binding affinity change (%) in HeLa/TFEB cells treated or not with CeO2 nanoparticles or taxol for 24 h (Figure 3A).
FLOW Human
Show All 27 Publications.

Reviews for Annexin V Kit (NBP2-29373) (2) 42

Average Rating: 4
(Based on 2 reviews)

Reviews using NBP2-29373:
Filter by Applications
ICC/IF
(1)
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Images Ratings Applications Species Date Details
Immunocytochemistry/Immunofluorescence Annexin V NBP2-29373
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4
reviewed by:
Anonymous
ICC/IF 02/20/2020
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Summary

ApplicationImmunocytochemistry/Immunofluorescence
Lot092519

Comments

CommentsSpecies: Human
Sample type: Cell culture
Assay time: 1-2 hours
Followed instructions: Yes
Serum Annexin V NBP2-29373
Enlarge
4
reviewed by:
Anonymous
02/20/2020
View

Summary

ApplicationSerum
Lot092519

Comments

CommentsSpecies: Human
Sample type: Cell culture
Assay time: 1-2 hours
Followed instructions: Yes

Product General Protocols

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ICC/IF Video Protocol

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FITC NBP2-29373

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Anonymous
02/20/2020
Application: ICC/IF
Species:

Anonymous
02/20/2020
Application:
Species:

Bioinformatics

Gene Symbol ANXA5