Anti-Mouse CD16.2 (Clone 9E9) In Vivo Antibody

Product Name:	Anti-Mouse CD16.2 (Clone 9E9) In Vivo Antibody - Ultra Low Endotoxin
Product Code:	IVMB0299
Size:	1 mg, 5 mg, 25 mg, 50 mg, 100 mg
Clone:	9E9
Protein:	CD16.2
Product Type:	Monoclonal Antibody
Synonyms:	FcgammaRIV
Isotype:	IgG
Reactivity:	Mouse
Applications:	B, FC, In Vivo
Formulation:	This monoclonal antibody is aseptically packaged and formulated in 0.01 M phosphate buffered saline (150 mM NaCl) PBS pH 7.2 - 7.4 with no carrier protein, potassium, calcium or preservatives added. Due to inherent biochemical properties of antibodies, certain products may be prone to precipitation over time. Precipitation may be removed by aseptic centrifugation and/or filtration.
Endotoxin Level:	<0.5 EU/mg as determined by the LAL method
Purity:	≥98% monomer by analytical SEC ⋅ >95% by SDS Page
Product Preparation:	Functional grade preclinical antibodies are manufactured in an animal free facility using in vitro cell culture techniques and are purified by a multi-step process including the use of protein A or G to assure extremely low levels of endotoxins, leachable protein A or aggregates.

Applications:	B, FC, In Vivo
Reactivity:	Mouse

Host Species:	Armenian Hamster
Specificity:	9E9 activity is primarily directed against mouse CD16.2 / FcgammaRIV but can also bind and block FcgammaRIII in vivo.
Antigen Distribution:	FcgammaRIV is expressed on the cell membrane of splenic and bone marrow dendritic cells, monocytes, and macrophages as well as peripheral blood monocytes, neutrophils, thioglycollate-elicited macrophages, and myeloid cells. FcgammaRIV is absent from lymphoid populations, T cells, B cells, NK cells, and other granulocytes.
Concentration:	≥ 5.0 mg/ml
Endotoxin Level:	<0.5 EU/mg as determined by the LAL method
Formulation:	This monoclonal antibody is aseptically packaged and formulated in 0.01 M phosphate buffered saline (150 mM NaCl) PBS pH 7.2 - 7.4 with no carrier protein, potassium, calcium or preservatives added. Due to inherent biochemical properties of antibodies, certain products may be prone to precipitation over time. Precipitation may be removed by aseptic centrifugation and/or filtration.
Purity:	≥98% monomer by analytical SEC ⋅ >95% by SDS Page
Preparation:	Functional grade preclinical antibodies are manufactured in an animal free facility using in vitro cell culture techniques and are purified by a multi-step process including the use of protein A or G to assure extremely low levels of endotoxins, leachable protein A or aggregates.
Storage and Handling:	Functional grade preclinical antibodies may be stored sterile as received at 2-8°C for up to one month. For longer term storage, aseptically aliquot in working volumes without diluting and store at -80°C. Avoid Repeated Freeze Thaw Cycles.

Fcgamma receptors are the primary mediators of IgG effector responses, and individual Fc receptors (FcR) have different affinities for different IgG subclasses1. Four FcgammaRs are present in mice2, and FcgammaRIV (FcgammaRL3, CD16.2) binds to IgG2a, IgG2b3, and IgE4, but not IgG1 or IgG33. FcgammaRIV is a high-affinity receptor for monomeric IgG2a and IgG2b and a low-affinity IgE receptor for both IgEa and IgEb, binding to aggregates but not monomers4. Additionally, IgE immune complexes can displace IgG2 from FcgammaRIV. Surface expression of FcgammaRIV requires gamma chain coexpression in vitro and in vivo3. FcgammaRIV and the gamma chain are upregulated on bone marrow-derived monocytes by IFN-gamma and LPS and are downregulated by TGF-β and IL-4.

According to surface plasmon resonance, 9E9 has strong reactivity to FcgammaRIV as well as low level binding to FcgammaRII and FcgammaRIII2. In vivo, 9E9 binds and blocks FcgammaRIII only when 9E9 first binds FcgammaRIV on the same effector cell, resulting in concurrent inhibition of FcgammaRIII and FcgammaRIV. Native 9E9 binds to FcgammaRII and FcgammaRIII via the Fc.

9E9 was produced by immunizing Armenian hamsters with an FcgammaRIV-IgG1 fusion protein consisting of the extracellular domain of FcgammaRIV fused to a mouse IgG1 Fc portion (D265A-variant deficient in Fc-receptor binding)3. Splenic B cells were then fused to a mouse fusion partner, and hybridoma clones were screened for binding to CHO-K1-FcgammaRIV cells expressing FcgammaRIV.

Blocking studies with 9E9 show that FcgammaRIV is necessary for IgG2a and IgG2b mediated platelet clearance in vivo1. Additionally, blocking FcgammaRIV with 9E9 reduces B-cell depletion2. 9E9 also interferes with immune complex binding to FcgammaRIV3 and can block FcgammaRIII on macrophages and neutrophils2.

Protein:	CD16.2
Research Area:	Immunology
Pathogen Testing:	To protect mouse colonies from infection by pathogens and to assure that experimental preclinical data is not affected by such pathogens, all of Assay Genie's Purified Functional Antibodies are tested and guaranteed to be negative for all pathogens in the IDEXX IMPACT I Mouse Profile.
Shipping:	2-8°C