General information
SALSA
® MLPA
® Probemix P110 FCGR mix 1 and SALSA
® MLPA
® Probemix P111 FCGR mix 2 are
research use only (RUO) assays for the detection of genomic rearrangements and point mutations in the 180 kb FCGR2/3 locus at 1q23.3.
Receptors for the Fc portion of IgG play an essential role in the protection of the organism against foreign antigens by removing antigen-antibody complexes from the circulation. Receptors are present on monocytes, macrophages, neutrophils, natural killer (NK) cells, and T and B lymphocytes. The receptors participate in diverse functions, such as phagocytosis of immune complexes and modulation of antibody production by B cells. Genes for several low-affinity Fc gamma receptors are clustered on chromosome 1q23.3 Within a 180 kb chromosomal area are genes for the FCGR2A, FCGR2B, FCGR2C, FCGR3A, and FCGR3B proteins. In addition, this region contains genes for the HSPA6 and HSPA7 heat shock proteins.
Due to high similarity between these
FCGR genes and their close proximity, gene rearrangements are frequent in this chromosomal region. Various functionally relevant polymorphisms (SNPs) in these genes, as well as copy number variation of the
FCGR2C,
FCGR3A, and
FCGR3B genes, have been reported. The MLPA probemixes P110 FCGR mix 1 and P111 FCGR mix 2 cover the mentioned
FCGR genes and are intended to detect both copy number changes of these genes as well as frequent polymorphisms and point mutations, such as
FCGR2B -386G/C,
FCGR2B -120A/T,
FCGR3A p.Val158Phe,
FCGR2B p.Ile232Thr,
FCGR2A p.His166Arg, and
FCGR2A p.Gln62Trp. Probes for
FCGR3B Human Neutrophil Antigen 1 (HNA1) alleles NA1, NA2 and SH, and
FCGR2C STOP, classic and non-classic ORF haplotypes are also included.
These products are not CE/FDA registered for use in diagnostic procedures. The SALSA® MLPA® technique is covered by US patent 6,955,901 and corresponding patents outside the US. The purchase of these products includes a license to use only this amount of product solely for the purchaser’s own use.
Probemix content
P110-C1 FCGR mix 1 contains 40 MLPA probes with amplification products between 130 and 494 nucleotides (nt). P111-C1 FCGR mix 2 contains 38 MLPA probes with amplification products between 130 and 490 nucleotides (nt).
Both P110 FCGR mix 1 and P111 FCGR mix 2 contain 14 probes each for determination of copy number changes and genomic rearrangements. Furthermore, P110 FCGR mix 1 also contains 18 mutation-specific probes and P111 FCGR mix 2 contains 16 mutation-specific probes which will only generate a signal when the mutation is present. In addition, for both probemixes eight reference probes are included that detect autosomal chromosomal locations. Complete probe sequences and the identity of the genes detected by the reference probes are available online (
www.mrcholland.com).
These probemixes contain nine quality control fragments generating amplification products between 64 and 105 nt: four DNA Quantity fragments (Q-fragments), two DNA Denaturation fragments (D-fragments), one Benchmark fragment, and one chromosome X and one chromosome Y-specific fragment. More information on how to interpret observations on these control fragments can be found in the MLPA General Protocol and online at
www.mrcholland.com.
SALSA® Reference Selection & Binning DNA SD038
The selection of suitable reference DNA samples that can be used with P110 FCGR mix 1 and P111 FCGR mix 2 is complicated. To facilitate the selection of suitable reference DNA samples from your own sample collection, a reference selection DNA sample is provided with these probemixes from MRC Holland. SALSA
® Reference Selection & Binning DNA SD038 should only be used for initial experiments on DNA samples from healthy individuals with the intention to identify suitable reference samples.
SD038 should not be used as a reference sample in subsequent experiments.
In addition, SALSA
® Reference Selection & Binning DNA SD038 can also be used for binning of all mutation-specific probes that are included in P110 FCGR mix 1 and P111 FCGR mix 2 probemixes. Inclusion of one reaction of SD038 in initial MLPA experiments is essential as it can be used to aid in data binning of the peak pattern using Coffalyser.Net. Furthermore, binning DNA should be included in the experiment whenever changes have been applied to the set-up of the capillary electrophoresis device (e.g. when capillaries have been renewed). SD038 should never be used as a reference sample in the MLPA data analysis, neither should it be used in quantification of mutation signals. For further details, please consult the SD038 product description, available online:
www.mrcholland.com.
These products are for research use only (RUO).
Sample DNA
Sample DNA developed for this product: