Preparation, labelling and quality control of 99mTc-HMPAO-labelled WBCs had to follow the procedures described in the SmPC of Ceretec™.
Sample preparation, labeling, post-staining, and detection procedures were the same as in single labeling experiments, except that rabbit anti-OsLRR1 serum and mouse anti-OsHIR1 serum (both 1 50 in 1% PBS-BSA) were applied simultaneously to the sample grid to detect the target proteins.
Commercially available MLT preparation labeled to contain 3 mg MLT was purchased from the local market.
There are several commercial sets of control probes made to measure the hybridization quality, as well as the RNA sample preparation, labelling and fragmentation process [28].
The cDNA preparation, labelling, and Affymetrix ATH-121501 GeneChip® hybridization was carried out by AGRF (Australian Genome Research Facility, http://www.agrf.org.au) using 15 ug of total RNA and three biological replicates.
When introduced early in the sample preparation, label-based methods are preferred for analyses that require extensive prefractionation/enrichment approaches, as lack of reproducibility of the above methods can be compensated for by a unique processing of the mixed sample.
For the replicate experiments, cell cultures, ChIP sample preparation, labeling and hybridization were performed independently.
Microarray sample preparation, labeling, and array hybridizations were performed as described in the Supporting Information.
Experimental details of tissue RNA preparation, labelling of cRNA, and microarray hybridization were as described in Methods.
DK conducted RNA preparation, labelling and microarray hybridization and together with GN conducted the statistical evaluation of raw data.
The RNA quality and quantity assessment, probe preparation, labeling, hybridization and image scan were carried out in the CCHMC Affymetrix Core using standard procedure.
