Description:
Receptortyrosinekinases(RTKs)playimportantrolesinconvertingextracellulartointracellularsignals.Theyareactivatedbythebindingofspecificgrowthfactors,cytokines,orhormones,whichresultsinautophosphorylation.TyrosineautophosphorylationgeneratesrecruitmentsitesfordownstreamsignalingproteinscontainingSrchomology-2(SH2).BindingofdifferentSH2domain-containingsignalingproteinsrepresentstheactivationofdifferentsignalingpathways.Therefore,theactivationofsignalingpathwayscanbeassessedthroughexaminationofSH2domainbinding.SignosisprovidedSH2Domain-BasedRTKProfilingPlateArray,whichallowsassessingtheactivationofRTKsignalingpathwaysthroughtheexaminationof46SH2domainbindingsimultaneously.ApplicableGrid:
ListofArrayedSH2Domains
| 1 | 2 | 3 | 4 | 5 | 6 | 1 | 2 | 3 | 4 | 5 | 6 | |
| A | 3BP2FP-0001 | CHN1FP-0015 | GRB10FP-0030 | LNKFP-0044 | P1K3R1-D2 | RaLPFP-0067 | 3BP2FP-0001 | CHN1FP-0015 | GRB10FP-0030 | LNKFP-0044 | P1K3R1-D2 | RaLPFP-0067 |
| B | ALXFP-0004 | CHN2FP-0016 | GRB14FP-0031 | LYNFP-0045 | P1K3R2-D2 | RIN1FP-0068 | ALXFP-0004 | CHN2FP-0016 | GRB14FP-0031 | LYNFP-0045 | P1K3R2-D2 | RIN1FP-0068 |
| C | BCAR3FP-0006 | CRKFP-0018 | GRB2FP-0032 | MATKFP-0046 | PLCG1-D1FP-0058 | RASA1 | BCAR3FP-0006 | CRKFP-0018 | GRB2FP-0032 | MATKFP-0046 | PLCG1-D1FP-0058 | RASA1 |
| D | BLKFP-0007 | CRKLFP-0019 | HCKFP-0036 | MISTFP-0047 | PLCG2-D2FP-0061 | SH2D2A | BLKFP-0007 | CRKLFP-0019 | HCKFP-0036 | MISTFP-0047 | PLCG2-D2FP-0061 | SH2D2A |
| E | BMX-3FP-0009 | CTENFP-0021 | ITKFP-0038 | NCK1FP-0048 | PTPN6-D1FP-0065 | SH2D4A | BMX-3FP-0009 | CTENFP-0021 | ITKFP-0038 | NCK1FP-0048 | PTPN6-D1FP-0065 | SH2D4A |
| F | BRDG1FP-0010 | FESFP-0025 | JAK1FP-0039 | NCK2FP-0049 | PTPN6-D2FP-0066 | SHCFP-0078 | BRDG1FP-0010 | FESFP-0025 | JAK1FP-0039 | NCK2FP-0049 | PTPN6-D2FP-0066 | SHCFP-0078 |
| G | BTKFP-0011 | FYNFP-0028 | LCKFP-0042 | NSP1FP-0050 | PTPNII-D1FP-0063 | GST | BTKFP-0011 | FYNFP-0028 | LCKFP-0042 | NSP1FP-0050 | PTPNII-D1FP-0063 | GST |
| H | CBLCFP-0014 | GRAPFP-0029 | LCP2FP-0043 | P1K3R1-D1 | PTPNII-D2FP-0064 | Blank | CBLCFP-0014 | GRAPFP-0029 | LCP2FP-0043 | P1K3R1-D1 | PTPNII-D2FP-0064 | Blank |
Principle:
SH2domain-basedRTKprofilingPlateArrayisanELISA-likeassay. SH2domainsare utilizedfor immobilizationonthemicrotiterwellsandananti-phosphotyrosine antibodyalongwithstreptavidinconjugatedtohorserADIshperoxidase(HRP)fordetection. Total46differentSH2domainsarecoatedindifferentwellsand46bindingscanbemeasured. Witha96wellmicroplate,twosamplescanbecomparedamongthebindings. Thetestsampleisallowedtoreactsimultaneouslywiththe domainandtheantibody.Afterincubation,thewellsarewashedtoremoveunbound-labeledantibody.AHRPsubstrate,TMB,isaddedtoresultinthedevelopmentofabluecolor.ThecolordevelopmentisthenstoppedwiththeadditionofStopSolutionchangingthecolortoyellow.Absorbanceismeasuredspectrophotometricallyat450nm.
Data:
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