Product Name Catalog # Price   Qty
SH2 Domain-Based RTK Profiling Plate Array DA-0001 $519
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Characterization

Description:

Receptor tyrosine kinases (RTKs) play important roles in converting extracellular to intracellular signals.They are activated by the binding of specific growth factors, cytokines, or hormones, which results in autophosphorylation. Tyrosine autophosphorylation generates recruitment sites for downstream signaling proteins containing Src homology-2 (SH2). Binding of different SH2 domain-containing signaling proteins represents the activation of different signaling pathways. Therefore, the activation of signaling pathways can be assessed through examination of SH2 domain binding. Signosis provided SH2 Domain-Based RTK Profiling Plate Array, which allows assessing the activation of RTK signaling pathways through the examination of 46 SH2 domain binding simultaneously.

 
Benefits:
  • Multiplex - A single assay allows the measurement of the interaction of RTK with 46 SH2 domains.
  • Quantitative comparison - The difference of individual proteins in the interaction of the SH2 domains can be quantitatively compared and analyzed.
  • Simple - The assay is performed the same as the procedure of ELISA.

Applicable Grid:

List of Arrayed SH2 Domains

  1 2 3 4 5 6 1 2 3 4 5 6
A 3BP2 FP-0001 CHN1 FP-0015 GRB10 FP-0030 LNK FP-0044 P1K3R1-D2 RaLP FP-0067 3BP2 FP-0001 CHN1 FP-0015 GRB10 FP-0030 LNK FP-0044 P1K3R1-D2 RaLP FP-0067
B ALX FP-0004 CHN2 FP-0016 GRB14 FP-0031 LYN FP-0045 P1K3R2-D2 RIN1 FP-0068 ALX FP-0004 CHN2 FP-0016 GRB14 FP-0031 LYN FP-0045 P1K3R2-D2 RIN1 FP-0068
C BCAR3 FP-0006 CRK FP-0018 GRB2 FP-0032 MATK FP-0046 PLCG1-D1 FP-0058 RASA1 BCAR3 FP-0006 CRK FP-0018 GRB2 FP-0032 MATK FP-0046 PLCG1-D1 FP-0058 RASA1
D BLK FP-0007 CRKL FP-0019 HCK FP-0036 MIST FP-0047 PLCG2-D2 FP-0061 SH2D2A BLK FP-0007 CRKL FP-0019 HCK FP-0036 MIST FP-0047 PLCG2-D2 FP-0061 SH2D2A
E BMX-3 FP-0009 CTEN FP-0021 ITK FP-0038 NCK1 FP-0048 PTPN6-D1 FP-0065 SH2D4A BMX-3 FP-0009 CTEN FP-0021 ITK FP-0038 NCK1 FP-0048 PTPN6-D1 FP-0065 SH2D4A
F BRDG1 FP-0010 FES FP-0025 JAK1 FP-0039 NCK2 FP-0049 PTPN6-D2 FP-0066 SHC FP-0078 BRDG1 FP-0010 FES FP-0025 JAK1 FP-0039 NCK2 FP-0049 PTPN6-D2 FP-0066 SHC FP-0078
G BTK FP-0011 FYN FP-0028 LCK FP-0042 NSP1 FP-0050 PTPNII-D1 FP-0063 GST BTK FP-0011 FYN FP-0028 LCK FP-0042 NSP1 FP-0050 PTPNII-D1 FP-0063 GST
H CBLC FP-0014 GRAP FP-0029 LCP2 FP-0043 P1K3R1-D1 PTPNII-D2 FP-0064 Blank CBLC FP-0014 GRAP FP-0029 LCP2 FP-0043 P1K3R1-D1 PTPNII-D2 FP-0064 Blank

Principle

SH2 domain-based RTK profiling Plate Array is an ELISA-like assay.   SH2 domains are  utilized for  immobilization on the microtiter wells and an anti-phosphotyrosine  antibody along with streptavidin conjugated to horseradish peroxidase (HRP) for detection.  Total 46 different SH2 domains are coated in different wells and 46 bindings can be measured.  With a 96 well microplate, two samples can be compared among the bindings.   The test sample is allowed to react simultaneously with the  domain and the antibody. After incubation, the wells are washed to remove unbound-labeled antibody. A HRP substrate, TMB, is added to result in the development of a blue color. The color development is then stopped with the addition of Stop Solution changing the color to yellow. Absorbance is measured spectrophotometrically at 450 nm.


Data


Analysis RTK activation with SH2-based RTK profiling assay. HeLa cells were treated with or without EGF 100ng/ml for 5 minutes. The cell lysates were prepared and subjected to the assay.

 
 

Literature

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