CD200/OX2 Antibody [Unconjugated]

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 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.

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.

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 (//pubmed.ncbi.nlm.nih.gov/31790427), licensed under a CC-BY license. Not internally tested by R&D Systems.

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 (//pubmed.ncbi.nlm.nih.gov/22642833), licensed under a CC-BY license. Not internally tested by R&D Systems.

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 (//pubmed.ncbi.nlm.nih.gov/31790427), licensed under a CC-BY license. Not internally tested by R&D Systems.

CD200R1 and CD200 protein levels in the post-mortem brain of Parkinson’s disease patients. Expression of CD200R1 protein isoforms 1 and 4 and CD200 proteins in the substantia nigra (SN) (a), frontal cortex (b), and hippocampus (c) tissue homogenates from control individuals (C, n = 4–8) and Parkinson’s disease patients (PD, n = 11–21) by western blot. Representative immunoblots are presented. Protein levels were normalized relative to beta tubulin. Data are expressed as fold change relative to C and are depicted as individual points with bars showing means ± SEM. *p < 0.05 and **p < 0.01 vs. C; Mann-Whitney test. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/35296683), licensed under a CC-BY license. Not internally tested by R&D Systems.

CD200R1 and CD200 protein levels in the post-mortem brain of Parkinson’s disease patients. Expression of CD200R1 protein isoforms 1 and 4 and CD200 proteins in the substantia nigra (SN) (a), frontal cortex (b), and hippocampus (c) tissue homogenates from control individuals (C, n = 4–8) and Parkinson’s disease patients (PD, n = 11–21) by western blot. Representative immunoblots are presented. Protein levels were normalized relative to beta tubulin. Data are expressed as fold change relative to C and are depicted as individual points with bars showing means ± SEM. *p < 0.05 and **p < 0.01 vs. C; Mann-Whitney test. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/35296683), licensed under a CC-BY license. Not internally tested by R&D Systems.

CD200R1 and CD200 protein levels in the post-mortem brain of Parkinson’s disease patients. Expression of CD200R1 protein isoforms 1 and 4 and CD200 proteins in the substantia nigra (SN) (a), frontal cortex (b), and hippocampus (c) tissue homogenates from control individuals (C, n = 4–8) and Parkinson’s disease patients (PD, n = 11–21) by western blot. Representative immunoblots are presented. Protein levels were normalized relative to beta tubulin. Data are expressed as fold change relative to C and are depicted as individual points with bars showing means ± SEM. *p < 0.05 and **p < 0.01 vs. C; Mann-Whitney test. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/35296683), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Mu, Rt
• Applications: WB, Simple Western, IHC
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

CD200/OX2 Antibody [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant mouse CD200
  Gln31-Gly232
  Accession # O54901
• Specificity: Detects mouse CD200 in direct ELISAs and mouse CD200 and rat CD200 Western blots. In direct ELISAs, approximately 50% cross-reactivity with recombinant human CD200 is observed.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: CD200
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Immunohistochemistry 5-15 ug/mL
  • Simple Western 2.5 ug/mL
  • Western Blot 0.25 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for CD200/OX2 Antibody [Unconjugated]

• antigen identified by monoclonal MRC OX-2
• CD200 antigenMOX1
• CD200 molecule
• CD200
• MOX1
• MOX2
• MOX2MRC
• MRC OX-2 antigen
• MRC
• OX-2 membrane glycoprotein
• OX-2

Background

CD200, also known as OX-2, is a 45 kDa type I transmembrane immunoregulatory protein that belongs to the immunoglobulin superfamily (1, 2). The mouse CD200 cDNA encodes a 278 amino acid (aa) precursor that includes a 30 aa signal sequence, a 202 aa extracellular domain (ECD), a 27 aa transmembrane segment, and a 19 aa cytoplasmic domain. The ECD is composed of one Ig-likeV-type and one Ig-like C2-type domain (3). Splice variants of CD200 have been described in human but not in mouse. Within the ECD, mouse CD200 shares 76% and 94% aa sequence identity with human and rat CD200, respectively. CD200 is widely but not ubiquitously expressed (4). Its receptor (CD200R) is restricted primarily to mast cells, basophils, macrophages, and dendritic cells, which suggests myeloid cell regulation as the major function of CD200 (5‑7). CD200 knockout mice are characterized by increased macrophage number and activation, and are predisposed to autoimmune disorders (8). CD200 and CD200 R associate via their respective N-terminal Ig-like domains (9). In myeloid cells, CD200 R initiates inhibitory signals following receptor-ligand contact (6, 7, 10). In T cells, CD200 functions as a costimulatory molecule that is independent of the CD28 pathway (11). Several additional CD200 R-like molecules have been identified in human and mouse, but their capacity to interact with CD200 is controversial (12, 13). Several viruses encode CD200 homologs which are expressed on infected cells during the lytic phase (14, 15). Like CD200 itself, viral CD200 homologs also suppress myeloid cell activity, enabling increased viral propagation (5, 14‑16).

1. Gorczynski, R.M. (2005) Curr. Opin. Invest. Drugs 6:483.
2. Barclay, A.N. et al. (2002) Trends Immunol. 23:285.
3. Chen, Z. et al. (1997) Biochim. Biophys. Acta 1362:6.
4. Wright, G.J. et al. (2001) Immunology 102:173.
5. Shiratori, I. et al. (2005) J. Immunol. 175:4441.
6. Cherwinski, H.M. et al. (2005) J. Immunol. 174:1348.
7. Fallarino, F. et al. (2004) J. Immunol. 173:3748.
8. Hoek, R.M. et al. (2000) Science 290:1768.
9. Hatherley, D. and A.N. Barclay (2004) Eur. J. Immunol. 34:1688.
10. Jenmalm, M.C. et al. (2006) J. Immunol. 176:191.
11. Borriello, F. et al. (1997) J. Immunol. 158:4548.
12. Gorczynski, R. et al. (2004) J. Immunol. 172:7744.
13. Hatherley, D. et al. (2005) J. Immunol. 175:2469.
14. Foster-Cuevas, M. et al. (2004) J. Virol. 78:7667.
15. Cameron, C.M. et al. (2005) J. Virol. 79:6052.
16. Langlais, C.L. et al. (2006) J. Virol. 80:3098.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for CD200/OX2 Antibody (AF3355)(5)

Publications using AF3355

• A Comella Bo, T Valente, A Miguez, V Brito, S Gines, C Solà, M Straccia, JM Canals CD200 is up-regulated in R6/1 transgenic mouse model of Huntington&#039;s disease PLoS ONE, 2019-12-02;14(12):e0224901. 2019-12-02 [PMID: 31790427] (IHC-Fr, Western Blot, Mouse)
• Ting‐Xin Jiang, Hans I‐Chen Harn, Kuang‐Ling Ou, Mingxing Lei, Cheng‐Ming Chuong Comparative regenerative biology of spiny ( Acomys cahirinus) and laboratory ( Mus musculus) mouse skin Experimental Dermatology 2019-04-01 [PMID: 30734959]
• Lyons A, Downer E, Costello D, Murphy N, Lynch M Dok2 mediates the CD200Fc attenuation of Abeta-induced changes in glia. J Neuroinflammation, 2012-05-29;9(0):107. 2012-05-29 [PMID: 22642833] (Western Blot, Mouse)
• Garza LA, Yang CC, Zhao T, Blatt HB, Lee M, He H, Stanton DC, Carrasco L, Spiegel JH, Tobias JW, Cotsarelis G Bald scalp in men with androgenetic alopecia retains hair follicle stem cells but lacks CD200-rich and CD34-positive hair follicle progenitor cells. J. Clin. Invest., 2011-01-04;121(2):613-22. 2011-01-04 [PMID: 21206086] (IHC-P, Mouse)
• Gorczynski RM, Chen Z, Shivagnahnam S Potent immunosuppression by a bivalent molecule binding to CD200R and TGF-betaR. Transplantation, 2010-07-27;90(2):150-9. 2010-07-27 [PMID: 20548263] (Western Blot, Mouse)

Bioinformatics

• Gene Symbol: CD200
• Uniprot:
  • O54901(Goat)