Product Description |
Recombinant CHO-K1 cells
stably overexpress low affinity immunoglobulin
gamma Fc region receptor III-A
(FcγRIIIa / CD16A 158F) 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 binding
affinity and stability of the Fc region of antibodies, Fc fusion proteins, or
antibody based biologics that bind with CD16A 158F. GenScript also offers a stable cell line expressing
the CD16A of 158V allotype (Cat. No. M00597) 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 |
2 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) (Cat. No.21127-022,
Gibco), 10% FBS (Cat. No.10099-141, Gibco), 8 μg/ml Puromycin (Cat. No.A11138-03,
Gibco), 400μg/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. It can
be used to isolate populations of these cells by antibodies directed towards
CD16, 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
higher affinity 158 V/V homodimer of CD16A is present on approximately 20% of
the population and the lower affinity F/F and F/V heterodimers 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 CD16A 158F on CHO-K1/CD16A 158F cells.
The CHO-K1/CD16A 158F cells (blue) and the negative control CHO-K1 cells (red) were stained with PerCP/Cyanine5.5 anti-human CD16 Antibody (Cat. No. 302028, Biolegend).
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