Rabbit anti cd11b

SeeTable S3. Tumor microvascular density was assessed using the rat anti-mouse CD31 (PECAM-1, clone SZ31) antibody (Dianova). The following antibodies were used to characterize immune cell subpopulations in the LL/2 tumors: rat anti-CD45 (Novus Biologicals), rabbit anti-CD11 b, rat anti-Ly6G/Ly6c (Novusbio), mouse anti-CD3, rat anti-CD4, goat anti-CD8 (Santa Cruz).

Western blot was performed as previously described (Tramullas et al., 2010 (link)). Whole-cell lysates were prepared from the lumbosacral region of the spinal cord and total protein was determined using the Quan-iT protein assay kit (Invitrogen, Ireland). Equal amounts of protein were subjected to electrophoresis on 4–12% gradient gels (NuPAGE, Invitrogen, Ireland) and transferred to a polyvinylidene difluoride membrane (Bio Rad, Ireland). Membranes were then incubated with goat anti-GFAP (1:1000; Sigma, Ireland), the astrocyte marker, rabbit anti-CD11b (1:500; Novus Biologicals, UK), a microglia activation marker; and mouse anti β-actin (1:15000; Sigma, Ireland). Immunoreactivity was detected with Pierce ECL detection reagent (Thermo Scientific, IL, USA) and visualized using a luminescent image analyzer (LAS-3000, Fujifilm, Ireland). The relative density of the immunoreactive bands was quantified using Fiji Is Just ImageJ (FIJI) software (Schindelin et al., 2012 (link)) and normalized to the relative density of mouse anti-β actin (Sigma, Ireland) for each sample. No significant differences were observed in the protein expression levels of β-Actin between groups.