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Cell Biology Apoptosis Annexin V Reagents
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Annexin V-PE-Cy5 Apoptosis Staining / Detection Kit (ab14159)

  • Datasheet
  • SDS
  • Protocol Booklet
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Key features and details

  • Assay type: Direct
  • Platform: Flow cytometer, Fluorescence microscope
  • Assay time: 10 min
  • Sample type: Adherent cells, Suspension cells

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Overview

  • Product name

    Annexin V-PE-Cy5 Apoptosis Staining / Detection Kit
    See all Annexin V/ANXA5 kits
  • Sample type

    Adherent cells, Suspension cells
  • Assay type

    Direct
  • Assay time

    0h 10m
  • Product overview

    Annexin V-PE-Cy5 Apoptosis Staining / Detection Kit (ab14159) 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 be easily 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 for fixation. Results can be analyzed by flow cytometry or fluorescence microscopy.
    Visit our FAQs page for tips and troubleshooting.

  • Platform

    Flow cytometer, Fluorescence microscope

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 100 tests
    Annexin V-PE 1 x 500µl
    Binding Buffer 1 x 50ml
  • Research areas

    • Cell Biology
    • Apoptosis
    • Annexin V Reagents
    • Cancer
    • Invasion/microenvironment
    • Apoptosis
    • Detection kits
    • Annexin V
    • Cardiovascular
    • Atherosclerosis
    • Apoptosis
    • Kits/ Lysates/ Other
    • Kits
    • ELISA Kits
    • ELISA Kits
    • Apoptosis marker and proteins ELISA kits
    • Kits/ Lysates/ Other
    • Kits
    • Apoptosis Kits
    • Annexin V Assays
    • Annexin V-PE-Cy5
    • Cancer
    • Cell Death
    • Apoptosis
    • Apoptosis Markers
    • Annexin V
    • Annexin V Detection Kits
    • Cancer
    • Cell Death
    • Apoptosis
    • Apoptosis Markers
    • Annexin V
    • Annexin V Reagents
  • Function

    This protein is an anticoagulant protein that acts as an indirect inhibitor of the thromboplastin-specific complex, which is involved in the blood coagulation cascade.
  • Involvement in disease

    Pregnancy loss, recurrent, 3
  • Sequence similarities

    Belongs to the annexin family.
    Contains 4 annexin repeats.
  • Domain

    The [IL]-x-C-x-x-[DE] motif is a proposed target motif for cysteine S-nitrosylation mediated by the iNOS-S100A8/A9 transnitrosylase complex.
    A pair of annexin repeats may form one binding site for calcium and phospholipid.
  • Post-translational
    modifications

    S-nitrosylation is induced by interferon-gamma and oxidatively-modified low-densitity lipoprotein (LDL(ox)) possibly implicating the iNOS-S100A8/9 transnitrosylase complex.
  • Target information above from: UniProt accession P08758 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Alternative names

    • Anchorin CII
    • Annexin 5
    • Annexin A5
    • Annexin V
    • Annexin-5
    • Annexin5
    • AnnexinA5
    • AnnexinV
    • ANX 5
    • ANX A5
    • ANX5
    • ANXA5
    • ANXA5_HUMAN
    • Calphobindin I
    • CBP-I
    • Endonexin II
    • ENX 2
    • ENX2
    • Lipocortin V
    • PAP I
    • PAP-I
    • Placental anticoagulant protein 4
    • Placental anticoagulant protein I
    • PLACENTAL PROTEIN 4
    • PP 4
    • Pp4
    • RPRGL3
    • Thromboplastin inhibitor
    • VAC-alpha
    • Vascular anticoagulant-alpha
    see all

Associated products

  • Related Products

    • FCCP, mitochondrial oxidative phosphorylation uncoupler (ab120081)
    • Cycloheximide, Protein synthesis inhibitor (ab120093)
    • SB431542, ALK inhibitor (ab120163)
    • Z-VAD(OH)-FMK, Irreversible general caspase inhibitor (ab120382)
    • Z-VAD(OMe)-FMK, Cell permeable, irreversible pan-caspase inhibitor (ab120487)
    • Doxorubicin hydrochloride, Topoisomerase II inhibitor (ab120629)
    • Dorsomorphin (Compound C), AMP-kinase inhibitor (ab120843)
    • Annexin V/ANXA5-FITC Apoptosis Detection Reagent (500X) (ab14082)
    • Propidium Iodide (ab14083)
    • 10X Annexin V/ANXA5 Binding Buffer (ab14084)
    • Annexin V-FITC Apoptosis Staining / Detection Kit (ab14085)
    • Annexin V-Cy3 Apoptosis Staining / Detection Kit (ab14142)
    • Calcein AM, fluorescent dye for cell viability (ab141420)
    • Annexin V/ANXA5-Cy3 Apoptosis Staining / Detection Reagent (ab14143)
    • Annexin V-Cy3 Apoptosis Staining / Detection Kit with SYTOX (ab14144)
    • Annexin V/ANXA5-Cy5 Apoptosis Staining / Detection Reagent (ab14147)
    • Annexin V-Cy5 Apoptosis Staining / Detection Kit (ab14150)
    • Annexin V-EGFP Apoptosis Staining / Detection Kit (ab14153)
    • Annexin V/ANXA5-PE Apoptosis Staining / Detection Reagent (ab14154)
    • Annexin V-PE Apoptosis Staining / Detection Kit (ab14155)
    • Annexin V/ANXA5-Biotin Apoptosis Staining / Detection Reagent (ab14165)

Protocols

  • Protocol Booklet

Click here to view the general protocols

Datasheets and documents

    • Datasheet
    • SDS
  • References (0)

    Publishing research using ab14159? Please let us know so that we can cite the reference in this datasheet.

    ab14159 has not yet been referenced specifically in any publications.

    Customer reviews and Q&As

    Show All Reviews Q&A
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    1-2 of 2 Abreviews or Q&A

    Question

    Thank you for prompt reply. I am sorry for incorrect enquiry, but I would ask you about possibility to use ab14159Annexin V-PE-Cy5 Apoptosis Detection Kit for detection apoptoticcells expressing GFP.

    Read More

    Abcam community

    Verified customer

    Asked on Feb 20 2012

    Answer



    We do not have any experimentaldata for theuse of this kitwith GFP, howeverI think this should be ok sincethe kit uses the tandem (PE-Cy5) for which the excitation wavelengthis 488 nm and the emission 670nm. This should not interfere with the GFP wavelengths of 395 nm (Ex)and at 470 nm (Em).

    I hope this is helpful.

    Read More

    Abcam Scientific Support

    Answered on Feb 20 2012

    Question

    BATCH NUMBER 67413 ORDER NUMBER 76761 DESCRIPTION OF THE PROBLEM 1. Binding Buffer was not included in the kit. 2. When replaced by a buffer containing kit, antibody did not bind. There were no positive signal in the FACS analysis when using the positive (apoptotic cells) control. 3. No signals were detected in the experimental groups also. SAMPLE HEK 293 cells with or without serum withdrawal induced apoptosis for analysis in FACS PRIMARY ANTIBODY Annexin V-PG-Cy5 SECONDARY ANTIBODY No DETECTION METHOD FACS POSITIVE AND NEGATIVE CONTROLS USED 293 Cells with or without serum withdrawal induced apoptosis SAMPLE PREPARATION Cells were trypsined and washed in pbs for two times and then washed one time in Annexin binding buffer, then annexin V antibody was added, FACS were performed in 15 min. TRANSFER AND BLOCKING CONDITIONS Annexin V binding buffer, incubation 15 time RT HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? three HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes WHAT STEPS HAVE YOU ALTERED? Different dilutions of antibody had been tried and different positive control cells ( ok cells with ATP depletion, which has been shown previously induced apoptosis, therefore annexin V positive cells. ADDITIONAL NOTES The volume of the primary antibody (Annexin V-PG-Cys) is also less than what is indicated in the information sheet. Only about 50 -100 ul in the tube.

    Read More

    Abcam community

    Verified customer

    Asked on Apr 25 2005

    Answer

    I'm very sorry to hear you are having problems with this kit and for the shortages you have experienced. I have arranged for 2 kits to be sent to you as soon as possible (order reference 80894) and apologise for the inconvenience. I also enclose the detailed protocol to help you, Please let me know if you need anything else and if you are satisfied with the new kits. Introduction: The Annexin V-PE-Cy5 Apoptosis Detection Kit 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 be easily 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 for fixation. Results can be analyzed by flow cytometry or fluorescence microscopy. II. Annexin V-PE-Cy5 Assay Protocol: A. Incubation of cells with Annexin V-PE-Cy5 1. Induce apoptosis by desired method. 2. Collect 1-5 x 105 cells by centrifugation. 3. Resuspend cells in 500 µl of 1X Binding Buffer. 4. Add 5 µl of Annexin V-PE-Cy5. 5. Incubate at room temperature for 5 min in the dark. Proceed to B or C below depending on method of analysis. B. Quantification by Flow Cytometry Analyze Annexin V-PE-Cy5 binding by flow cytometry (Ex = 488 nm; Em = 670 nm). For analyzing adherent cells, gently trypsinize and wash cells once with serum-containing media before incubation with Annexin V-PE-Cy5 (A.3-5). C. Detection by Fluorescence Microscopy 1. Place the cell suspension from Step A.5 on a glass slide. Cover the cells with a glass coverslip. For analyzing adherent cells, grow cells directly on a coverslip. Following incubation (A.5), invert coverslip on glass slide and visualize cells. The cells can also be washed and fixed in 2% formaldehyde before visualization. (Cells must be incubated with Annexin V-PE-Cy5 before fixation since any cell membrane disruption can cause nonspecific binding of Annexin V to PS on the inner surface of the cell membrane.) 2. Observe the cells under a fluorescence microscope using the PE-Cy5 filter. Cells which have bound Annexin V-PE-Cy5 will show bright purple staining in the plasma membrane. III. Storage and stability: Store kit at 4oC.

    Read More

    Abcam Scientific Support

    Answered on Apr 27 2005

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