Product Description |
Recombinant CHO-K1 cells
stably express low affinity immunoglobulin
gamma Fc region receptor III-A
(FcγRIIIa / CD16A 158V) complex on
the cell surface, including 1 alpha chain and 2 gamma chains. The surface expression
of CD16A is validated by FACS
analysis. This stable cell line product is designed for measuring the binding
affinity and stability of the Fc region of antibodies, Fc fusion proteins, or antibody
based biologics that bind with CD16A
158V. GenScript also offers a stable cell line expressing the CD16A of
158F allotype (Cat. No. M00598) for FcγRIIIa
polymorphism study. |
Applications |
Cell-based binding assay; use as immunogen for antibody development |
Expressed Gene |
Codon
optimized from NM_000569.6 (for FCGR3A); codon
optimized from NM_004106.2 (for FCER1G); no expressed tags |
Host Cell |
CHO-K1 |
Quantity |
Two
vials of frozen cells (>1×106 per
vial in 1 ml) |
Storage |
Store cells in liquid nitrogen immediately upon
receipt. Thaw and recover cells within one year from the date received. |
Mycoplasma Status |
Negative. The mycoplasma test was performed with
MycoAlert™ PLUS Mycoplasma Detection Kit (Cat. No. LT07-318, Lonza). |
Culture Properties |
Adherent |
Freeze Medium |
95% Culture Medium, 5%
(V/V) DMSO (Cat. No. D2650, Sigma) |
Complete Growth Medium |
Ham’s F-12K (Kaighn’s)
(Cat. No. 21127-022, Gibco), 10% FBS (Cat. No. 10099-141, Gibco) |
Culture Medium |
Ham’s F-12K (Kaighn’s), 10%
FBS, 8 μg/ml Puromycin (Cat. No. A11138-03, Gibco), 200μg/ml Hygromycin B (Cat.
No. 10687010, Invitrogen) |
Gene Synonyms |
FCGR3A, FCG3, FCGR3 (for Fc Fragment of IgG Receptor
IIIa); FCER1G, FCRG (for Fc fragment of IgE receptor Ig) |
Background |
CD16 is a
cluster of differentiation molecule found on the surface of natural killer
cells, monocytes and macrophages. Antibodies against CD16 can be used to
isolate populations of these cells using fluorescent-activated cell sorting or
magnetic-activated cell sorting. CD16 has been identified as Fc receptors
FcγRIIIa (CD16A) and FcγRIIIb (CD16B). The two isoforms of the CD16A
stimulatory receptor found on natural killer (NK) cells and macrophages differ
by one amino acid at codon 158 (V/F) and are important for antibody-dependent
cell-mediated cytotoxicity (ADCC). The high affinity 158 V/V homodimer of CD16a
is present on approximately 20% of the population and the lower affinity F/F
homodimer and F/V heterodimer are present in the remaining 80%. Human FcγRIIIA (CD16A)
is a low-affinity receptor for the Fc portion of IgG. It is a protein complex
composed of an alpha chain encoded by FCGR3A (Fc Fragment of IgG Receptor IIIa)
and two gamma chains encoded by FCER1G (Fc Fragment of IgE Receptor Ig). |
Figure 1: FACS analysis of cell surface expression of human CD16A 158V on CHO-K cells.
CHO-K1/CD16A 158V cells (blue) and negative control CHO-K1 cells (red) were stained with PE-Cy™5 Mouse Anti-Human CD16 antibody (Cat. No. 555408, BD Biosescience).
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