{"Equipment":"<strong>Reagents and equipment for sample preparation</strong><br />\r\n<ul>\r\n<li>Spleen Dissociation Kit (Miltenyi Biotec)</li>\r\n<li>gentleMACS Dissociator (Miltenyi Biotec)</li>\r\n<li>gentleMACS C Tubes (Miltenyi Biotec)</li>\r\n<li>FACS buffer: 2% Fetal Bovine Serum, 10mM HEPES, 2mM EDTA in HBSS without Ca2+ and Mg2+</li>\r\n<li>ACK Lysis buffer: 150mM NH4Cl, 10mM KHCO3, 0.1mM Na2EDTA in H2O</li>\r\n<li>CellTrics filters 100&mu;m (Sysmex)</li>\r\n<li>BD Horizon Brilliant Stain Buffer Plus</li>\r\n<li>Anti-CD16/CD32 antibody - Fc block (BD Biosciences)</li>\r\n</ul>","Procedure":"<strong>Sample preparation</strong><br />\r\n<ul>\r\n<li>Dissociate spleens enzymatically in gentleMACS C Tubes according to the Spleen Dissociation Kit protocol.</li>\r\n<li>Fill the tubes with cell suspensions up to 10 mL with FACS buffer and transfer the suspensions through 100 &mu;m cell strainer into 15mL tubes.</li>\r\n<li>Centrifuge samples at 400 g for 8 minutes at 4&deg;C.</li>\r\n<li>Resuspend pellets in 1 mL of ACK Lysis buffer and incubate for 3 minutes at room temperature.</li>\r\n<li>Add FACS buffer up to 5 mL and filter cell suspension through the 100 &mu;m cell strainer.</li>\r\n<li>Count total number of live cells using Trypan Blue stain and Cellometer Auto T4 with Nexcelom cell counting chambers.</li>\r\n<li>Transfer 2.5x106 of live cells per well into a 96-well U bottom plate. Centrifuge &nbsp;plate at 400 g for 4 minutes at 4&deg;C.</li>\r\n<li>Discard supernatant and proceed with staining.</li>\r\n</ul>\r\n<strong>Staining and data acquisition</strong><br />\r\n<ul>\r\n<li>Resuspend pellets in 50 &mu;L of staining cocktail (antibodies including Fc block (1:200) in FACS buffer containing 20% of Brilliant stain buffer):</li>\r\n<li>\r\n<table style=\"border-collapse: collapse; width: 100.027%; height: 733.15px; border-color: #000000; margin-left: auto; margin-right: auto;\" border=\"2\" cellpadding=\"0cm 5.4pt\">\r\n<tbody>\r\n<tr style=\"height: 34.1px;\">\r\n<td style=\"width: 16.132%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle; background-color: #c2e0f4;\"><strong>Specificity</strong></td>\r\n<td style=\"width: 18.7087%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle; background-color: #c2e0f4;\"><strong>Fluorochrome</strong></td>\r\n<td style=\"width: 17.7004%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle; background-color: #c2e0f4;\"><strong>Clone</strong></td>\r\n<td style=\"width: 14.4516%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle; background-color: #c2e0f4;\"><strong>Catalog #</strong></td>\r\n<td style=\"width: 21.1689%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle; background-color: #c2e0f4;\"><strong>Manufacturer</strong></td>\r\n<td style=\"width: 11.8794%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle; background-color: #c2e0f4;\"><strong>Dilution</strong></td>\r\n</tr>\r\n<tr style=\"height: 34.1px;\">\r\n<td style=\"width: 16.132%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">CD8a&nbsp;</td>\r\n<td style=\"width: 18.7087%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">BUV395</td>\r\n<td style=\"width: 17.7004%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">53-6.7</td>\r\n<td style=\"width: 14.4516%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">563786</td>\r\n<td style=\"width: 21.1689%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">BD Biosciences</td>\r\n<td style=\"width: 11.8794%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">1:200</td>\r\n</tr>\r\n<tr style=\"height: 34.1px;\">\r\n<td style=\"width: 16.132%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">CD4</td>\r\n<td style=\"width: 18.7087%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">BUV496</td>\r\n<td style=\"width: 17.7004%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">RM4-5</td>\r\n<td style=\"width: 14.4516%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">741050</td>\r\n<td style=\"width: 21.1689%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">BD Biosciences</td>\r\n<td style=\"width: 11.8794%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">1:400</td>\r\n</tr>\r\n<tr style=\"height: 34.1px;\">\r\n<td style=\"width: 16.132%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">F4/80</td>\r\n<td style=\"width: 18.7087%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">BUV563</td>\r\n<td style=\"width: 17.7004%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">T45-2342</td>\r\n<td style=\"width: 14.4516%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">749284</td>\r\n<td style=\"width: 21.1689%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">BD Biosciences</td>\r\n<td style=\"width: 11.8794%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">1:400</td>\r\n</tr>\r\n<tr style=\"height: 34.1px;\">\r\n<td style=\"width: 16.132%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">CD19</td>\r\n<td style=\"width: 18.7087%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">BUV615</td>\r\n<td style=\"width: 17.7004%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">1D3</td>\r\n<td style=\"width: 14.4516%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">751213</td>\r\n<td style=\"width: 21.1689%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">BD Biosciences</td>\r\n<td style=\"width: 11.8794%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">1:800</td>\r\n</tr>\r\n<tr style=\"height: 34.1px;\">\r\n<td style=\"width: 16.132%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">&gamma;&delta;TCR</td>\r\n<td style=\"width: 18.7087%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">BUV661</td>\r\n<td style=\"width: 17.7004%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">GL3</td>\r\n<td style=\"width: 14.4516%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">750410</td>\r\n<td style=\"width: 21.1689%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">BD Biosciences</td>\r\n<td style=\"width: 11.8794%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">1:200</td>\r\n</tr>\r\n<tr style=\"height: 34.1px;\">\r\n<td style=\"width: 16.132%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">CD21/CD35</td>\r\n<td style=\"width: 18.7087%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">BUV737</td>\r\n<td style=\"width: 17.7004%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">7G6</td>\r\n<td style=\"width: 14.4516%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">612810</td>\r\n<td style=\"width: 21.1689%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">BD Biosciences</td>\r\n<td style=\"width: 11.8794%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">1:2000</td>\r\n</tr>\r\n<tr style=\"height: 34.1px;\">\r\n<td style=\"width: 16.132%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">CD44</td>\r\n<td style=\"width: 18.7087%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">BUV805</td>\r\n<td style=\"width: 17.7004%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">IM7</td>\r\n<td style=\"width: 14.4516%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">741921</td>\r\n<td style=\"width: 21.1689%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">BD Biosciences</td>\r\n<td style=\"width: 11.8794%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">1:400</td>\r\n</tr>\r\n<tr style=\"height: 34.1px;\">\r\n<td style=\"width: 16.132%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">CD5&nbsp;</td>\r\n<td style=\"width: 18.7087%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">BV421</td>\r\n<td style=\"width: 17.7004%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">53-7.3</td>\r\n<td style=\"width: 14.4516%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">562739</td>\r\n<td style=\"width: 21.1689%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">BD Biosciences</td>\r\n<td style=\"width: 11.8794%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">1:400</td>\r\n</tr>\r\n<tr style=\"height: 17.05px;\">\r\n<td style=\"width: 16.132%; height: 17.05px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">Viability</td>\r\n<td style=\"width: 18.7087%; height: 17.05px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">SYTOX Blue</td>\r\n<td style=\"width: 17.7004%; height: 17.05px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">-</td>\r\n<td style=\"width: 14.4516%; height: 17.05px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">S34857</td>\r\n<td style=\"width: 21.1689%; height: 17.05px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">Invitrogen</td>\r\n<td style=\"width: 11.8794%; height: 17.05px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">1:20000</td>\r\n</tr>\r\n<tr style=\"height: 17.05px;\">\r\n<td style=\"width: 16.132%; height: 17.05px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">CD11c&nbsp;</td>\r\n<td style=\"width: 18.7087%; height: 17.05px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">BV605</td>\r\n<td style=\"width: 17.7004%; height: 17.05px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">N418</td>\r\n<td style=\"width: 14.4516%; height: 17.05px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">117334</td>\r\n<td style=\"width: 21.1689%; height: 17.05px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">BioLegend</td>\r\n<td style=\"width: 11.8794%; height: 17.05px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">1:400</td>\r\n</tr>\r\n<tr style=\"height: 34.1px;\">\r\n<td style=\"width: 16.132%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">Ly6G&nbsp;</td>\r\n<td style=\"width: 18.7087%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">BV650</td>\r\n<td style=\"width: 17.7004%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">1A8</td>\r\n<td style=\"width: 14.4516%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">740554</td>\r\n<td style=\"width: 21.1689%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">BD Biosciences</td>\r\n<td style=\"width: 11.8794%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">1:400</td>\r\n</tr>\r\n<tr style=\"height: 34.1px;\">\r\n<td style=\"width: 16.132%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">BST2 (CD317)</td>\r\n<td style=\"width: 18.7087%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">BV711</td>\r\n<td style=\"width: 17.7004%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">927</td>\r\n<td style=\"width: 14.4516%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">747604</td>\r\n<td style=\"width: 21.1689%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">BD Biosciences</td>\r\n<td style=\"width: 11.8794%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">1:400</td>\r\n</tr>\r\n<tr style=\"height: 34.1px;\">\r\n<td style=\"width: 16.132%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">Ly6C&nbsp;</td>\r\n<td style=\"width: 18.7087%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">FITC</td>\r\n<td style=\"width: 17.7004%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">AL-21</td>\r\n<td style=\"width: 14.4516%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">553104</td>\r\n<td style=\"width: 21.1689%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">BD Biosciences</td>\r\n<td style=\"width: 11.8794%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">1:800</td>\r\n</tr>\r\n<tr style=\"height: 34.1px;\">\r\n<td style=\"width: 16.132%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">MHCII&nbsp;</td>\r\n<td style=\"width: 18.7087%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">BB630</td>\r\n<td style=\"width: 17.7004%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">M5/114.15.2</td>\r\n<td style=\"width: 14.4516%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">624294</td>\r\n<td style=\"width: 21.1689%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">BD Biosciences</td>\r\n<td style=\"width: 11.8794%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">1:800</td>\r\n</tr>\r\n<tr style=\"height: 34.1px;\">\r\n<td style=\"width: 16.132%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">KLRG1</td>\r\n<td style=\"width: 18.7087%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">BB700</td>\r\n<td style=\"width: 17.7004%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">2F1</td>\r\n<td style=\"width: 14.4516%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">742199</td>\r\n<td style=\"width: 21.1689%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">BD Biosciences</td>\r\n<td style=\"width: 11.8794%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">1:400</td>\r\n</tr>\r\n<tr style=\"height: 34.1px;\">\r\n<td style=\"width: 16.132%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">CD49d</td>\r\n<td style=\"width: 18.7087%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">BB790</td>\r\n<td style=\"width: 17.7004%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">R1-2</td>\r\n<td style=\"width: 14.4516%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">624296</td>\r\n<td style=\"width: 21.1689%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">BD Biosciences</td>\r\n<td style=\"width: 11.8794%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">1:400</td>\r\n</tr>\r\n<tr style=\"height: 17.05px;\">\r\n<td style=\"width: 16.132%; height: 17.05px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">CD23&nbsp;</td>\r\n<td style=\"width: 18.7087%; height: 17.05px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">PE</td>\r\n<td style=\"width: 17.7004%; height: 17.05px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">B3B4</td>\r\n<td style=\"width: 14.4516%; height: 17.05px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">101607</td>\r\n<td style=\"width: 21.1689%; height: 17.05px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">BioLegend</td>\r\n<td style=\"width: 11.8794%; height: 17.05px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">1:800</td>\r\n</tr>\r\n<tr style=\"height: 34.1px;\">\r\n<td style=\"width: 16.132%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">CD161</td>\r\n<td style=\"width: 18.7087%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">PE-CF594</td>\r\n<td style=\"width: 17.7004%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">PK136</td>\r\n<td style=\"width: 14.4516%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">562864</td>\r\n<td style=\"width: 21.1689%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">BD Biosciences</td>\r\n<td style=\"width: 11.8794%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">1:800</td>\r\n</tr>\r\n<tr style=\"height: 34.1px;\">\r\n<td style=\"width: 16.132%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">CD25</td>\r\n<td style=\"width: 18.7087%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">PE-Cy7</td>\r\n<td style=\"width: 17.7004%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">PC61</td>\r\n<td style=\"width: 14.4516%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">552880</td>\r\n<td style=\"width: 21.1689%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">BD Biosciences</td>\r\n<td style=\"width: 11.8794%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">1:400</td>\r\n</tr>\r\n<tr style=\"height: 34.1px;\">\r\n<td style=\"width: 16.132%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">GITR (CD357)</td>\r\n<td style=\"width: 18.7087%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">APC</td>\r\n<td style=\"width: 17.7004%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">DTA-1</td>\r\n<td style=\"width: 14.4516%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">624346</td>\r\n<td style=\"width: 21.1689%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">BD Biosciences</td>\r\n<td style=\"width: 11.8794%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">1:800</td>\r\n</tr>\r\n<tr style=\"height: 34.1px;\">\r\n<td style=\"width: 16.132%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">CD11b&nbsp;</td>\r\n<td style=\"width: 18.7087%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">APC-R700</td>\r\n<td style=\"width: 17.7004%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">M1/70</td>\r\n<td style=\"width: 14.4516%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">564985</td>\r\n<td style=\"width: 21.1689%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">BD Biosciences</td>\r\n<td style=\"width: 11.8794%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">1:2000</td>\r\n</tr>\r\n<tr style=\"height: 34.1px;\">\r\n<td style=\"width: 16.132%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">CD62L&nbsp;</td>\r\n<td style=\"width: 18.7087%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">APC-Cy7</td>\r\n<td style=\"width: 17.7004%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">MEL-14</td>\r\n<td style=\"width: 14.4516%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">560514</td>\r\n<td style=\"width: 21.1689%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">BD Biosciences</td>\r\n<td style=\"width: 11.8794%; height: 34.1px; border-color: #000000; border-style: solid; text-align: center; vertical-align: middle;\">1:2000</td>\r\n</tr>\r\n</tbody>\r\n</table>\r\n</li>\r\n</ul>\r\n<ul>\r\n<li>Incubate samples for 30 minutes in dark on ice.</li>\r\n<li>Wash samples with 200 &mu;L of FACS buffer and centrifuge for 4 minutes at 400 g at 4 &deg;C.</li>\r\n<li>Discard supernatant and resuspend pellets in 150 &mu;L of FACS buffer containing SYTOX Blue (diluted 1:40 000).</li>\r\n<li>Acquire 500 000 live cells.</li>\r\n</ul>","Purpose":"<p>This test differentiates immune cell sub-populations via flow cytometry in one panel.</p>","Experimental Design":"<ul>     <li><strong>Minimum number:</strong> 7M + 7F</li>     <li><strong>Age at test:</strong>  Week 16</li>     <li><strong>Sex:</strong> Both (sexually dimorphic)</li> </ul>","Notes":"<p><strong>Visual help for Gating</strong></p>\r\n<p><img src=\"https://api.mousephenotype.org/impress/image/9101b27c6fa0a48de267bf773abc9b1bc7fade39fde581f45ddf7e800a01df8b\"></p>"}