After three washes, streptavidin-ALP (Mabtech, Nacka Strand, Sweden) was added for 1 h at RT, followed by the addition of the substrate for spot development (Mabtech).
This may possibly result in local temperature runaway and/or hot spot development leading to several potentially unanticipated consequences for safety and integrity of the tube and hence the reactor.
Now, after decades of contention, spot development, ad-hoc rezoning efforts and fits-and-starts installation of infrastructure, the residents of what is commonly called South Richmond are being presented with a plan that many officials and community activists believe offers hope for more rational growth.
Plates were incubated for an additional 2 h at 37°C and spot development was done as described [16].
Plates were then incubated for 3 hours at 37°C for spot development, digitally photographed and counted manually using ImageJ software (National Institutes of Health, USA).
For both Phase I and II, protocols and reagents required for spot development were the centre's own.
Extravidin alkaline phosphatase conjugate (1 30,000 in PBS/1% BSA) and BCIP/NBT solution (bromochloroindolyle phophate/nitroblue tetrazolium; both from Sigma) were used for spot development.
Plates were incubated for an additional 2 h at 37 °C, and spot development was done as described in ref. Spot numbers were automatically determined with the use of a computer-assisted video image analyzer with KS ELISPOT 4.0 software (Carl Zeiss Vision).
All other protocol choices were left to the participants, including choices about: Elispot plate, antibodies, spot development, use of DNAse, resting of cells, T cell serum, cell counting and spot counting method.
NCLB severity was visually rated for each plot according to the lesion spot development in the middle-to-upper part leaves on a scale from 1 (susceptible) to 9 (resistant).
