Immunocytochemistry/ Immunofluorescence: Polarity Sensitive Indicator of Viability Apoptosis Microscopy Kit [NBP2-29382] - Quantitative analysis of pSIVA-IANBD™ fluorescence is rescued and NGF deprived neuronal ...read more
Immunocytochemistry/ Immunofluorescence: Polarity Sensitive Indicator of Viability Apoptosis Microscopy Kit [NBP2-29382] - Rescue of dying neurons: NGF withdrawal leads to cell death and pSIVA-IANBD(TM) fluorescence. ...read more
Immunocytochemistry/ Immunofluorescence: Polarity Sensitive Indicator of Viability Apoptosis Microscopy Kit [NBP2-29382] - Rescue of dying neurons. NGF withdrawal leads to cell death and pSIVA-IANBD(TM) fluorescence. ...read more
Immunofluorescence: Polarity Sensitive Indicator of Viability Apoptosis Microscopy Kit [NBP2-29382] - Rescue of dying photoreceptors in dystrophic rat neural retina. Image from verified customer review.
Hu, Mu, RtSpecies Glossary
ICC/IF, In vitro, LIM
Concentration is not relevant for this product. Please see the protocols for proper use of this product.
Polarity Sensitive Indicator of Viability Apoptosis Microscopy Kit Summary
Polarity Sensitive Indicatior of Viability (pSIVA (Annexin XII)) is an Annexin based, polarity sensitive probe for the spatiotemporal or kinetic analysis of apoptosis and other forms of cell death. pSIVA (Annexin XII) binding is reversible enabling researchers, for the first time, to detect transient PS exposure which is associated with normal physiological processes as well as with reversible or rescuable apoptosis cell death events. pSIVA (Annexin XII) is conjugated to IANBD, a polarity sensitive dye that fluoresces only when pSIVA is bound to the cell membrane. pSIVA's membrane-bound dependent fluorescence and reversible binding properties are a technological leap for detecting PS exposure and offer additional information on the apoptosis pathway and cell survival compared to Annexin V conjugates. Annexin V binding is nonreversible.
Apoptosis Microscopy Kit
Packaging, Storage & Formulations
Store at 4C. Do not freeze.
Concentration is not relevant for this product. Please see the protocols for proper use of this product.
In vitro assay
Live Imaging Microscopy
Please see PDF manual attached for specific usage information. Use in Immunocytochemistry/immunofluorescence reported in scientific literature (PMID 24162663)
Read 1 Review rated 5 using NBP2-29382 in the following applications:
Read Publications using NBP2-29382 in the following applications:
FAQs for Polarity Sensitive Indicator of Viability Kit (NBP2-29382). (Showing 1 - 10 of 18 FAQs).
What are the applications of pSIVA™-IANBD?
pSIVA™-IANBD is ideal for live imaging microscopy, confocal microscopy, fluorescence microscopy, in vivo applications and flow cytometry.
How does pSIVA™-IANBD work?
pSIVA™-IANBD binds to exposed phosphatidylserine (PS) residues on the surface of live cells.
How is pSIVA™-IANBD constructed?
pSIVA™ is a recombinant Annexin B12 construct, and is conjugated to IANBD which is a polar sensitive dye.
What are the similarities between Annexin V and pSIVA™-IANBD?
Both Annexin V and pSIVA™-IANBD bind to exposed PS on the surfaces of cells. Annexin V-FITC is the most commonly used Annexin V conjugate. Annexin V-FITC and pSIVA™-IANBD both excite at 488 nm and emit fluorescence at 530 nm.
What is the biological significance of exposed PS?
Exposed PS is a hallmark of early apoptosis and also occurs during other cell death processes. PS exposure may also occur during homeostasis and other normal cellular events.
What are the differences between Annexin V fluorescent conjugates and pSIVA™™™™-IANBD?
Annexin V fluorescent conjugates such as Annexin V-FITC fluoresce irrespective of whether or not they are bound to PS. In contrast pSIVA-IANBD fluoresces only when bound to PS. This is because pSIVA-IANBD is polar sensitive. pSIVA-IANBD fluoresces in non-polar but not polar environments. When pSIVA™-IANBD is bound to PS it is in the non-polar environment of the membrane lipid bilayer and fluoresces. However, when pSIVA-IANBD is not bound it is in the polar environment of the media or buffer and does not fluoresce. Annexin V-FITC requires washes to remove excess, unbound fluorescent molecules. However since pSIVA™-IANBD only fluoresces when bound to PS, washing is not needed.
What are the advantages of using pSIVA™™™-IANBD compared to Annexin V fluorescent conjugates?
Annexin V conjugates are primarily useful for flow cytometry. In contrast pSIVA™-IANBD is also ideal for live imaging microscopy, confocal microscopy, and immunofluorescence microscopy. pSIVA-IANBD, but not Annexin V-FITC can be used to study reversible or transient PS exposure events. This is because when PS exposure reverts to the inner bilayer, pSIVA-IANBD will be released into the polar environment of the media of buffer and no longer fluoresce. So why can’t you use Annexin V-FITC to study this phenomenon? If Annexin V-FITC were released back into the polar environment it would still fluorescence and hence it wouldn’t be possible to track transient PS exposure events because they wouldn’t be associated with loss of fluorescence as fluorescent conjugates like Annexin V-FITC are fluorescent irrespective of whether or not they are bound to their targets.
Why is pSIVA™™™-IANBD but not Annexin V-FITC useful for microscopy?
Annexin V-FITC has too much background for microscopy applications. It fluoresces all the time and washing is required to remove unbound Annexin V-FITC. In contrast since pSIVA-IANBD fluoresces only when bound to PS background is lacking. pSIVA-IANBD can be incubated with cells and monitored by live imaging because no wash is required.
Has pSIVA™-IANBD been published?
Yes, there are a number of pSIVA-IANBD publications. Please see the publication list in the pSIVA-IANBD manual.
Is pSIVA™ available in other conjugates besides IANBD?
No, pSIVA™ is not currently available in other conjugates.
Can pSIVA™-IANBD labeled cells be fixed?
Yes, once living cells are stained with pSIVA-IANBD they can be fixed and stained with other markers for flow cytometry or fluorescence microscopy.
Can be pSIVA™-IANBD be used in fixed tissue or frozen tissue?
No, pSIVA™-IANBD is used with live cells or living, fresh tissue.
Is pSIVA™-IANBD toxic?
pSIVA™-IANBD is not in of itself toxic to cells in the model systems heretofore described. However, since pSIVA™-IANBD binds to exposed PS, it may be possible that the binding up of exposed PS could have an unknown biological or biochemical effect in some model systems.
What species does pSIVA™-IANBD work in?
pSIVA-IANBD is not species specific and should theoretically works in any species ranging from worms to human. This pSIVA™-IANBD, like Annexin V-FITC, binds to exposed PS, which is a phenomenon from worms to humans. Theoretically, pSIVA™-IANBD should be suitable for any species that Annexin Vi-FITC has been used in.
How has pSIVA™-IANBD been used for in vivo applications?
An example would be injection into rat nerves to look at the effects on nerve injury (<a href="http://www.ncbi.nlm.nih.gov/pubmed/?term=Nature+Methods+7%3A67-73" target="_blank">Kim et al, Nature Methods 7:67-73, 2010</a>).
Where can I learn more about the nitty gritty behind pSIVA™-IANBD?
Please refer to the two 2010 development publications by Kim et al (<a href="http://www.ncbi.nlm.nih.gov/pubmed/?term=Nature+Methods+7%3A67-73" target="_blank">Nature Methods 7:67-73, 2010</a> Protocols 5:1`396-1405).
Our end user is working on tissues (epithelium of imaginal disk from drosophila leg). The tissues are kept growing on a culture medium and they manage to follow apoptosis properly with orange acrylin but not with annexin V because of background. Did you ever test any of your pSIVA apoptosis kits (NBP2-29382, NBP2-29611) on tissue culture instead of cell culture ? Is there a chance the proble will work on such an application ? Would you be able to provide a sample for this application ? As for the other samples, I'm still waiting on customer feedback and I'll keep you posted.
Yes, pSIVA has been used in tissue applications as denoted on the publication link. Specifically, I would encourage the research to look at the following publications from the link: 1. Diurnal, localized exposure of phosphatidylserine by rod outer segment tips in wild-type but not Itgb5-/- or Mfge8-/- mouse retina. Ruggiero L, MP Connor, J Chen, R Langen, SC Finnemann. PNAS 109:8145-4148 (2012). Live tissue imaging (mouse retina): Figs 4, 5. S4. pSIVA-IANBD was added to dissected live mouse retina and shown to label the tips of photoreceptor outer segments (POS). > The results suggested that phosphatidylserine (PS) exposure is specific to the POS surface. Furthermore, enhanced PS exposure preceded rod shedding and phagocytosis, suggesting that surface PS exposure promotes these processes. 2. Diurnal photoreceptor outer segment shedding: contributions of the RPE. Finnemann SC, L Ruggiero, MP Connor. Invest Opthalmol Vis Sci 53:4327 (2012). Live Imaging: mouse retina. pSIVA was used quantify the surface exposure of phosphotidylserine on the distal rod photoreceptor outer segments. 3. Characterization of cell cycle modifications induced by electric pulses in human corneal endothelium. Thi MH, Z He, N Campolmi, S Piselli, P Gain, M Peoc'H, JM Dumolllard, S Acquart, O Garraud, G Thuret. Acta Opthalmologica 90:s249 (2012). Live tissue imaging (human cornea organ cultures): pSIVA-IANBD was used to assess cell death following gene electrotransfer into corneal endothelial cells. > 3. Engineering a polarity-sensitive biosensor for time-lapse imaging of apoptotic processess and degeneration. Kim YE, J Chen, JR Chan, R Langen. Nature Methods 7:67-73 (2010a). Real-time live-cell imaging and time-lapse microscopy of apoptosis: Fig 2 (COS-7 cells), Fig 3 (rat neuronal degeneration), Fig 4 (rat axonal degeneration), Fig 5 (rescue of rat neuronal degeneration as visualized by pSIVA. (this was also used in a rat nerve injury tissue model) Please watch the pSIVA webinar on our website for additional information about pSIVA. The webinar is referred to March 25, 2014: Real-Time Tracking of Cell Death Webinar. We would certainly be pleased to offer any of your end user research customers a sample who is willing to provide a short explanation about what they are planning to do, their timeframe, and provide feedback about how it worked for them. For any samples, please specify if you
How many tests can you perform with this kit?
100 tests for the full sized kit and 25 tests for the 1 sample sized kit.