What kind of pipet tips can I use in ELISPOT assays?
For pipetting the cells, tips with wide openings are preferred to
reduce shear forces that induce apoptosis in T cells. (Avoid
touching the membrane with the pipet tips.)
I sometimes get streaks and irregular spots in the well bottoms. What could be causing this?
T cells can actively migrate during the assay’s duration. In particular, T cells that have been preactivated in vivo or in vitro can migrate rapidly, leaving a cytokine trail. Also, if the plates are shaken (due to banging of the incubator door, for example) or moved during the capture period, the cells will roll, leaving streaks and irregularly shaped spots.
Can I stack the plates in the incubator?
To ensure equal heat distribution, stacking the plates is not recommended.
What is the best way to dry the plates after development?
The best way is to put the plates upright into the flow hood over night.
Make sure not to dry them at higher temperatures than 37 °C, as the
membranes might crack.
How long can I store dried plates?
If kept protected from light, red plates are stable for up to five months.
Blue assay plates are more stable, but it would be advisable to keep them
protected from light as well.
Can I freeze T cells and still use them for Elispot?
If CTL protocols are followed and our freezing media are used,
human PBMCs can be frozen without any loss in function.
Please contact ctl-e@immunospot.com
for more information.
Does CTL provide assay protocols?
Yes, CTL provides protocols for single- and double-color ELISPOT assays.
CTL also offers wet laboratory training and assay consultation. Please
inquire at ctl-e@immunospot.com
for details.
Assay Performance
Scanning/ScAnalysis