Mouse/Rat CD200 Antibody

Detection of Mouse and Rat CD200 by Western Blot. Western blot shows lysates of mouse brain tissue and rat brain tissue. PVDF membrane was probed with 0.25 µg/mL of Goat Anti-Mouse/Rat CD200 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3355) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). A specific band was detected for CD200 at approximately 38-45 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

CD200 in Mouse Brain. CD200 was detected in perfusion fixed frozen sections of normal mouse brain using Goat Anti-Mouse/Rat CD200 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3355) at 15 µg/mL overnight at 4 °C. Tissue was stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to plasma membranes of hippocampal neurons. View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections.

Detection of Mouse and Rat CD200 by Simple Western^TM. Simple Western lane view shows lysates of mouse brain (cortex) tissue and rat brain (cortex) tissue, loaded at 0.2 mg/mL. A specific band was detected for CD200 at approximately 73-76 kDa (as indicated) using 2.5 µg/mL of Goat Anti-Mouse/Rat CD200 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3355) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Detection of Human CD200/OX2 by Western Blot Expression of CD200 protein isoforms in neocortex, hippocampus and striatum of HdhQ7/7 and HdhQ111/7 knock-in mice were analyzed at 6, 8 and 13 months of age by Western blot.CD200full and CD200tr isoforms are observed in the three brain regions. Quantification of CD200 total protein, both isoforms, have been performed (A) in neocortex at 6 (HdhQ7/7 = 13, HdhQ111/7 = 10), 8 (HdhQ7/7 = 7, HdhQ111/7 = 7) and 13 (HdhQ7/7 = 7, HdhQ111/7 = 7) months of age; (B) in hippocampus at 6 (HdhQ7/7 = 8, HdhQ111/7 = 13), 8 (HdhQ7/7 = 7, HdhQ111/7 = 7) and 13 (HdhQ7/7 = 7, HdhQ111/7 = 7) months of age; and (C) in striatum at 6 (HdhQ7/7 = 14, HdhQ111/7 = 10), 8 (HdhQ7/7 = 14, HdhQ111/7 = 10) and 13 (HdhQ7/7 = 7, HdhQ111/7 = 7) months of age. Bars represent the mean ± SEM. p-Values are the result of the multiple t-tests of the HdhQ111/7 genotype versus the respective HdhQ7/7 group. Representative Western blots for 8 months of age are shown as the percentage of immunosignal ratio between CD200 and beta -actin relative to respective HdhQ7/7 brain region. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31790427), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse CD200/OX2 by Western Blot A beta modulates CD200R-associated signaling. A. Incubation of purified astrocytes in the presence of A beta decreased CD200 expression as illustrated in the sample immunoblot and in the densitometric data (**P < 0.01; Student’s t-test for independent means; n = 6). B and C. A beta significantly increased CD200R mRNA on purified microglia (B) and the percentage of CD200R+ CD11b+ cells (C; *P < 0.05; **P < 0.01; Student’s t-test for independent means; n = 6). D and E. Whereas A beta decreased Dok2 phosphorylation (D), CD200Fc increased its phosphorylation in a time-dependent manner (E). Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/22642833), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human CD200/OX2 by Western Blot Expression of CD200 protein isoforms in R6/1 mouse brains at 8, 12, 20, and 30 weeks of age.(A) Neocortex, hippocampus and striatum were analyzed by Western blot and CD200 protein levels detected using beta -actin as loading control. Both isoforms, CD200full and the truncated CD200tr, are observed in all brain regions under analysis. Quantification of total CD200 protein (both isoforms) was measured in control (wt) and R6/1 mice: (B) in neocortex at 8 (wt = 6; R6/1 = 6), 12 (wt = 3; R6/1 = 3), 20 (wt = 3; R6/1 = 3) and 30 (wt = 3; R6/1 = 3) weeks of age; (C) in hippocampus at 8 (wt = 6; R6/1 = 6), 12 (wt = 5; R6/1 = 5), 20 (wt = 3; R6/1 = 3) and 30 (wt = 6; R6/1 = 6) weeks of age; and (D) in striatum at 8 (wt = 4; R6/1 = 5), 12 (wt = 3; R6/1 = 3), 20 (wt = 3; R6/1 = 3) and 30 (wt = 8; R6/1 = 8) weeks of age. Data are shown as percentage of immunosignal ratio between CD200 and beta -actin in R6/1 relative to their respective wild-type mice. Bars represent the means ± standard error of the means (SEM) of 3–8 animals. p-Values are the result of the multiple t-tests of the R6/1 genotype versus the respective wild-type group. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31790427), licensed under a CC-BY license. Not internally tested by R&D Systems.