Annexin V/ANXA5-FITC Apoptosis Detection Reagent (500X) (ab14082)

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.

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.

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.

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,

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.