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Annexin V Kit

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Annexin V Assay Kit - 100 Tests thumbnail image 1 Annexin V Assay Kit - 100 Tests thumbnail image 2
Annexin V Assay Kit - 100 Tests gallery image 1
Published customer image:
Annexin V:FITC Assay Kit used for cell cycle analysis of cells treated with Arsenic trioxide and Macuna macrocarpa extract by flow cytometry.
Image caption:
Annexin V-FITC/propidium iodide (PI) analyses of arsenic trioxide (ATO)- and/or crude methanolic extract of Mucuna macrocarpa (CMEMM)-treated cells. HL-60 (a) and Jurkat (b) cells (1 × 105 cells/mL) were first treated with 5 mM N-acetyl cysteine (NAC), 50 μM butylated hydroxytoluene (BHT), or 40 μM α-tocopherol (VitE) or untreated, followed by treatment with 0.1% DMSO (CTL), 2.5 μM ATO, and/or 50 μg/mL CMEMM as indicated for 24 h. Quantitative percentages of apoptotic cells of ATO/CMEMM-treated cells were measured by flow cytometry. Data represent the result from one of three independent experiments.

From: Lu KH, Lee HJ, Huang ML, Lai SC, Ho YL, Chang YS, Chi CW. Synergistic Apoptosis-Inducing Antileukemic Effects of Arsenic Trioxide and Mucuna macrocarpa Stem Extract in Human Leukemic Cells via a Reactive Oxygen Species-Dependent Mechanism.
Evid Based Complement Alternat Med. 2012;2012:921430.
Annexin V Assay Kit - 100 Tests gallery image 2
Jurkat cells were treated as shown with staurosporine at 1mM to induce apoptosis. The cells were then stained with annexin V FITC (ANNEX300F) and ReadiDrop™ propidium iodide (1351101). Apoptotic cells positive for Annexin V can be seen in the bottom right quadrant and dead cells positive for both Annexin and PI in the top right quadrant. Healthy cells are negative for both stains.

Annexin V:Biotin Assay Kit

Annexin V:FITC Assay Kit

Product Type
Kits
Product Code Applications Pack Size List Price Quantity
ANNEX100B
Datasheet
F
MSDS
100 Tests

ANNEX100F
Datasheet
F
MSDS
100 Tests

This test employs the property of Annexin V to bind to the membrane phospholipid phosphatidylserine (PS) in the presence of Ca2+. PS is exposed at the cell surface during the early stages of apoptosis. Detection of PS is a very sensitive method for detecting cells entering apoptosis, at a time point considerably ahead of nuclear changes such as DNA degradation.

The conjugation protocol used to prepare this product has not changed the native phospholipid binding properties of Annexin V. This protocol is designed to measure apoptosis easily and quickly in a sample of suspended cells.

View our complete list of formats and sizes of Annexin V kits

Product Details

Preservative Stabilisers
0.02%Sodium Azide (NaN3)
1%Bovine Serum Albumin
Preservative Stabilisers
0.02%Sodium Azide (NaN3)
1%Bovine Serum Albumin
Reagents in the Kit
Annexin V:Biotin1 x 0.5ml vial
Propidium Iodide1 x 1.6ml vial at 20ug/ml
Binding buffer1 x 50ml vial at x 4 concentrate


Note: This assay also requires streptavidin:FITC conjugate for visualisation (not supplied - see recommended useful reagents section).
Reagents in the Kit
Annexin V:FITC1 x 0.5ml vial
Propidium Iodide1 x 1.6ml vial at 20ug/ml
Binding buffer1 x 50ml vial at x 4 concentrate

Storage Information

Storage
Store at +4oC. DO NOT FREEZE.
This product should be stored undiluted. Should this product contain a precipitate we recommend microcentrifugation before use.
Storage
Store at +4oC. DO NOT FREEZE.
This product should be stored undiluted. This product is photosensitive and should be protected from light. Should this product contain a precipitate we recommend microcentrifugation before use.
Guarantee
Guaranteed until date of expiry. Please see product label.
Guarantee
Guaranteed until date of expiry. Please see product label.

More Information

UniProt
P08758
Entrez Gene
ANXA5
GO Terms
GO:0005509 calcium ion binding
GO:0007596 blood coagulation
GO:0004859 phospholipase inhibitor activity
GO:0007165 signal transduction
GO:0005544 calcium-dependent phospholipid binding
GO:0005737 cytoplasm
GO:0006916 anti-apoptosis
GO:0050819 negative regulation of coagulation
Regulatory
For research purposes only

Applications of Annexin V Kit

This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.
Application Name Verified Min Dilution Max Dilution
Flow Cytometry Neat
Flow Cytometry
Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using appropriate negative/positive controls.
Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using appropriate negative/positive controls.
Instructions For Use
1) Dilute the binding buffer 1:4 in distilled water (50ml binding buffer + 150ml distilled water).

2) Wash cells in PBS by gentle shaking or pipetting up and down.

3) Resuspend cells in 200ul pre-diluted binding buffer, adjusting to a cell density of 2-5 x 105 cells/ml.

4) Add 5ul Annexin V:Biotin to 195ul of the cell suspension prepared in step 3.

5) Mix and incubate for 15 minutes at room temperature.

6) Wash cells twice in 190ul of pre-diluted binding buffer.

7) Resuspend cells in 190ul pre-diluted binding buffer.

8) Add streptavidin:FITC conjugate.

9) Mix and incubate for 30 minutes in the dark, at room temperature.

10) Wash cells in 200ul pre-diluted binding buffer.

11) Resuspend cells in 190ul pre-diluted binding buffer.

12) Add 10ul of the Propidium Iodide solution.

13) Analyse by flow cytometry.

The flow cytometer is preferably set such that the Mean Fluorescence Intensity of the Annexin V negative population is between 1 and 10. Optimal parameter settings can be found using a positive control. For a positive control, incubate the cells with 3% formaldehyde in buffer during 30 minutes on ice. Wash away the formaldehyde and suspend the cells in cold binding buffer at 2-5 x 105 cells/ml. Proceed with step 2 as described above.
Instructions For Use
1) Dilute the binding buffer 1:4 in distilled water (50ml binding buffer +150ml distilled water)

2) Wash cells in PBS by gentle shaking or by pipetting up and down.

3) Resuspend cells in 200ul pre-diluted binding buffer, adjusting to a cell density of 2-5 x 105 cells/ml.

4) Add 5ul Annexin V:FITC to 195ul of the cell suspension prepared in step 3.

5) Mix and incubate for 10 minutes in the dark, at room temperature.

6) Wash cells in 200ul of pre-diluted binding buffer.

7) Resuspend cells in 190ul pre-diluted binding buffer.

8) Add 10ul of the Propidium Iodide solution.

9) Analyse by flow cytometry.

The flow cytometer is preferably set such that the Mean Fluorescence Intensity of the Annexin V negative population is between 1 and 10. Optimal parameter settings can be found using a positive control. For a positive control, incubate the cells with 3% formaldehyde in buffer during 30 minutes on ice. Wash away the formaldehyde and suspend the cells in cold binding buffer at 2-5 x 105 cells/ml. Proceed with step 2 as described above.
Copyright © 2020 Bio-Rad Antibodies (formerly AbD Serotec)

Useful Reagents Available

Description Product Code Applications Pack Size List Price Quantity
Streptavidin:FITC 710002 F 1 mg loader
Annexin V:FITC Assay Kit ANNEX100F F 100 Tests
Annexin V:FITC Assay Kit ANNEX300F F 300 Tests
Streptavidin:FITC STAR2B C F P 1 mg loader
Annexin V:Biotin Assay Kit ANNEX100B F 100 Tests
Annexin V:FITC Assay Kit ANNEX300F F 300 Tests

Application Based External Images

Flow Cytometry

Product Specific References

References for Annexin V Kit

  1. Lu, K.H. et al. (2010) In Vitro and In Vivo Apoptosis-Inducing Antileukemic Effects of Mucuna macrocarpa Stem Extract on HL-60 Human Leukemia Cells.
    Integr Cancer Ther. 9: 298-308.
  2. Yen, J.H. et al. (2010) Glycine tomentella Hayata inhibits IL-1β and IL-6 production, inhibits MMP-9 activity, and enhances RAW264.7 macrophage clearance of apoptotic cells.
    J Biomed Sci. 17: 83.
  3. Chen, C.W. et al. (2010) The signals of FGFs on the neurogenesis of embryonic stem cells.
    J Biomed Sci.17:33.
  4. Smith, K. et al. (2011) Mono- and tri-cationic porphyrin-monoclonal antibody conjugates: photodynamic activity and mechanism of action.
    Immunology. 132: 256-65.
  5. Lu, K.H. et al. (2012) Synergistic Apoptosis-Inducing Antileukemic Effects of Arsenic Trioxide and Mucuna macrocarpa Stem Extract in Human Leukemic Cells via a Reactive Oxygen Species-Dependent Mechanism.
    Evid Based Complement Alternat Med. 2012:921430.
  6. Koutsogiannaki, S. et al. (2015) Effects of cadmium and 17β-estradiol on Mytilus galloprovincialis redox status. Prooxidant-antioxidant balance (PAB) as a novel approach in biomonitoring of marine environments.
    Mar Environ Res. 103: 80-8.
  7. Lulia Irimie A et al. (2015) Inhibition of tumor necrosis factor alpha using RNA interference in oral squamous cell carcinoma.
    J. BUON. 20 (4): 1107-14.
  8. Hounkong, K. et al. (2015) Mechanisms of 1-hydroxy-2-hydroxymethylanthraquinone from Coptosapelta flavescens as an anti-giardial activity.
    Acta Trop. 146: 11-6.
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