Name
CD200 receptor-like 3, CD200R3, Mouse, mAb Ba103
Catalog nr
HM1103-1b (lot number and expiry date are indicated on the label)
Short description
The monoclonal antibody Ba103 recognizes mouse CD200 Receptor-like 3 (CD200R3), a protein of ~38 kDa or ~74 kDa (reduced or non-reduced condition, respectively). CD200R3 presumably exists as a disulfide-linked dimer on the cell surface, it is a single-pass type I membrane protein.
Size
1 mg
Application
FC , FS , IP , W
Technical datasheet
Description
The monoclonal antibody Ba103 recognizes mouse CD200 Receptor-like 3 (CD200R3), a protein of ~38 kDa or ~74 kDa (reduced or non-reduced condition, respectively). CD200R3 presumably exists as a disulfide-linked dimer on the cell surface, it is a single-pass type I membrane protein.
The OX-2, or CD200 receptor (CD200R), belongs to the immunoglobulin superfamily. CD200R contains two Ig-like extracellular domains, one Ig-like C2-type (immunoglobulin-like) domain and one Ig-like V-type (immunoglobulin-like) domain. and mediates inhibitory signals in myeloid cells. Mice deficient for the ligand CD200 develop enhanced experimental allergic encephalomyelitis and collagen-induced arthritis. Unlike CD200R, the CD200R-like receptors function as activating receptors. They contain short cytoplasmic tails and a lysine residue in the transmembrane region and are likely to signal via adaptor proteins such as DAP12, DAP10, FcR, or CD3.
CD200R3 is expressed in uterus and bone marrow-derived mast cells (at protein level). It is expressed in uterus, spleen, bone marrow-derived dendritic, basophil and mast cells. CD200R3 is also expressed in the lung of N. brasiliensis-infected mice. It is weakly expressed in brain, testis, lung and thymus.
CD200R3 is expressed preferentially on cells of the myeloid lineage, including mast cells and basophils. Both cell types are involved in the host defense system against pathogens and in the development of allergic disorders. They function as independent essential initiators of allergic reactions. Basophils are the least common leukocytes in the peripheral blood accounting approximately 0.5 % of all leukocytes. In vivo depletion of basophils by using monoclonal antibody Ba103 proved the essential role of basophils in the development and maintenance of IgE-mediated chronic allergic inflammation.
The activating CD200 receptors, like CD200R3, play an important role in IgE-independent mast cell and basophil activation. Monoclonal antibody Ba103 activates basophils ex vivo and elicits systemic anaphylaxis when administered in vivo. Cross-linking of CD200R3 on MC/9 cells by monoclonal antibody Ba103 induces degranulation. Furthermore, monoclonal antibody Ba103 is useful for depletion of basophils.
Cross Reactivity
 Cross reactant  Reactivity 
 CD200R  No
 CD200R2  No
 CD200R4  No
 CD200R5  No
Immunogen
Mouse primary basophils; cell-enriched bone marrow cells
Formulation
1 mg of 0.2 µm filtered antibody solution in PBS (exact concentration is indicated on the label).
Species
Rat IgG2b
Application

 

F

FC1,2

FS1-3

IA

IF

IP2

P

W2

Yes

 

 

 

 

No

 

 

 

 

 

 

 

 

N.D.

 

 

 

 

N.D.= Not Determined; F = Frozen sections; FC = Flow Cytometry; FS = Functional Studies; IA = Immuno Assays; IF = Immuno Fluorescence; IP = Immuno Precipitation; P = Paraffin sections; W = Western blot
Applications FC, FS and W were tested by Hycult Biotech.
Application notes
WB: reduced and non-reduced MC/9 lysate; band size 38kDa under reducing conditions and 74 kDa under nonreducing conditions
FC: Basophils and mast cells; no fixation needed
FS: antagonist in basophil-mediated allergic reactions; a single intravenous administration of 30 µg antibody . The biological activity of the antibody is drastic reduction in the basophile number in the peripheral blood and spleen, to approximately 20% of the normal level; also useful for mast cell degranulation; readout by measuring hexosaminidase activity
IP: mast cells were lysed with 1% NP-40 containing lysis buffer with proteinase inhibitor mixture before incubation with the antibody


Flow cytometric detection of MC/9 cells with monoclonal antibody to CD200R3.
Red line is only cells. Black line is isotype control (5 µg/ml). Green line is 5 µg/ml mAb Ba103.
Use
For flow cytometry, Western blotting and immuno precipitation, dilutions to be used depend on detection system applied. It is recommended that users test the reagent and determine their own optimal dilutions. The typical starting working dilution is 1:50. For functional studies, in vitro dilutions have to be optimized in user’s experimental setting.
Aliases
CD200 receptor-like 3, CD200RLb, cell surface glycoprotein OX-2 receptor 3
Positive control
MC/9
Negative control
CFTL-15 cells, T-cells, B-cells, macrophages, neutrophils
Storage and stability
Product should be stored at 4°C. Under recommended storage conditions, product is stable for one year.
References
1. Obata, K et al; Basophils are essential initiators of a novel type of chronic allergic inflammation. Blood 2007, 110: 913
2. Kojima, T et al; Mast cells and basophils are selectively activated in vitro and in vivo through CD200R3 in an IgE-independent manner. J Immunol 2007, 179: 7093
3. Khodoun, M et al; Peanuts can contribute to anaphylactic shock by activating complement. J Allergy Clin Immunol 2009, 123: 342
Precautions
For research use only. Not for use in or on humans or animals or for diagnostics. It is the responsibility of the user to comply with all local/state and federal rules in the use of this product. Hycult Biotech is not responsible for any patent infringements that might result from the use or derivation of this product.
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