TLSs were fixed in 10% formalin and stained using the Vectastain ABC Kit Vector Laboratoriess, USA), according to the manufacturer's instructions, in combination with rat anti-mouse CD31 (1 in 50 dilution) monoclonal antibody (BD Pharmingen, USA).
Polyclonal goat α-mouse podocalyxin (1∶50, R&D Systems) and monoclonal mouse α-pan cytokeratin antibodies (1∶50, Sigma) were used according to the manufacturer's instructions in combination with the appropriate secondary antibodies.
Two days later, T cell responses against the cognate peptides were analyzed by intracellular IFN-γ staining, using a cytokine staining kit (BD Pharmingen) according to the manufacturer's instructions in combination with goat anti-mouse CD4- and CD8-specific monoclonal antibody conjugates (BD Pharmingen).
Root canals were prepared using ProTaper instruments (Dentsply®, Konstanz, Germany) according to the manufacturer's instructions in combination with an Endo IT Professional (VDW®, Munich, Germany) at 250 rpm.
The supernatants were assayed in 10 mL samples for luciferase activity for 1 second using the Luciferase Reporter Assay system (Promega) according to the manufacturer's instructions, in combination with Wallac Victor 2 multilabel counter (PerkinElmer, Rodgau, Germany).
Total RNA from healthy specimens and 119 neoplastic specimens were extracted using Trizol® according to the manufacturer's instructions and in combination with Qiagen RNAeasy Mini Kit (Qiagen).
Spaced retrieval was offered in 11 studies used in combination with visual instructions, a stepwise approach, verbal instruction, and/or modeling.
Some studies have investigated the use of self-explanation in combination with direct instruction methods [ 65].
Most popular among them are glucosamine and chondroitin sulfate, often sold in combination tablets or capsules with instructions to take three a day.
RNA was extracted according to the manufacturer's instructions and purified using the RNeasy® mini kit in combination with DNAse treatment (RNAse-free DNAse set, Qiagen).
