Overview

Properties

Applications

Our Abpromise guarantee covers the use of ab14082 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
FM Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent dilution.
ICC/IF Use 1µl for 105 cells.

Images

  • Murine platelets were stimulated with CRP (5 µg/ml) in the presence of annexin V-fluorescein isothiocyanate for 10 minutes. The annexin V-positive population is indicated (AnV +ve).

References

This product has been referenced in:
  • Chen B  et al. Neuroprotection by chitosan nanoparticles in oxidative stress-mediated injury. BMC Res Notes 11:49 (2018). WB, IHC-P ; Human . Read more (PubMed: 29351805) »
  • Ghosh AS  et al. Hydrogen peroxide-induced apoptosis-like cell death in Entamoeba histolytica. Parasitol Int 59:166-72 (2010). Read more (PubMed: 20079879) »
See all 5 Publications for this product

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

Answer

Thank you for your inquiry.

I can confirm that ab14082 (5X Annexin V-FITC Apoptosis Detection Reagent) and ab14084 (10X Annexin V Binding Buffer) are the same reagents as in the kit ab14085(Annexin V-FITC Apoptosis Detection Kit).

Please note that ab14082 (5X Annexin V-FITC Apoptosis Detection Reagent) and ab14084 (10X Annexin V Binding Buffer) are more concentrated and need need to be diluted to 1x before using them.

I hope this information is helpful and wish you and your customer a good week.

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Answer

Thank you for your reply.

Both Annexin V-EGFP Apoptosis Detection Reagents (ab14152 and ab14082) contain a protocol booklet in their datasheets. These are the recommended protocols to use along with the Propidium Iodide reagent.

Please find below the link to these booklets:

https://www.abcam.com/ps/products/14/ab14082/documents/ab14082%20Annexin%20V%20FITC%20Apoptosis%20Detection%20Reagent%20(Website).pdf

https://www.abcam.com/ps/products/14/ab14152/documents/ab14152%20Annexin%20V%20EGFP%20Apoptosis%20Detection%20Reagent%20(Website).pdf

If you had any doubt or suggestions regarding the protocols or any other products, please do not hesitate to contact us back. We are more than happy to help.

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Answer

Thank you for your enquiry and your interest in our products.

The concentration of this product (ab14082) is 0.15 mg/ml, which is equivalent to 0.15 µg/ul. This is added to 500 µl makes it 0.15 µg/500 µl, which is 0.3 µg/ml. The PI is 1 mg/ml which is equivalent to 1 µg/µl, and in 500 µl it would make 2 µg/ml. (But note that the PI reagent actually has to be diluted to 250 µg/ml before use, which would mean that if this is done, the final concentration of PI in the 500 µl solution would be 0.5 µg/ml).

I hope this helps and if I can assist further, please do not hesitate to contact me.

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Answer

Thank you for contacting us for technical support, I'm sorry to hear you are experiencing problems with ab14082, I am trying to find out from the lab the function of the binding buffer and as soon as I find out I will forward to you this information. Here is the recommended protocol with this antibody, I will ask my colleagues to add it to the datasheet: A. Incubation of cells with Annexin V-FITC: 1. Induce apoptosis by desired methods. 2. Collect 1 x 105 cells by centrifugation. 3. Resuspend cells in 500 µl of 1X Annexin V Binding Buffer. 4. Add 1 µl of Annexin V-FITC and 1 µl Propidium Iodide. 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 cells by flow cytometry (Ex. = 488 nm; Em. = 530 nm) using FL1 channel for detecting Annexin V-FITC staining and FL2 channel for detecting PI staining. For adherent cells, trypsinize and gently wash cells with serum-containing medium before incubation with Annexin V-FITC. C. Detection by Fluorescence Microscopy: 1. Place the cell suspension on a glass slide, and cover with a glass coverslip. For analyzing adherent cells, grow cells directly on a coverslip. Following incubation, invert coverslip on a glass slide and visualize cells. The cells can also be washed with 1X Annexin V Binding Buffer and fixed in 2% formaldehyde before visualization. (Cells must be incubated with Annexin V-FITC before fixation because 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 a dual filter set for FITC and rhodamine, or separate filters. Cells that have bound Annexin V-FITC will show green staining on the plasma membrane. Cells that have lost membrane integrity will show red PI staining throughout the nuclei and a halo of green staining (FITC) on the plasma membrane. I hope this protocol will help you and will forward to you the information about the binding buffer as so as I receive it, Thank you for your patience,

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Answer

Thank you for your enquiry. The associated concentration for the Annexin V- FITC conjugate in the Annexin V- FITC Apoptosis Detection Reagent Kit is 250 ug/ml. Please contact us again if you require further assistance.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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