raybio human egfr phosphorylation antibody array 1 assay  (RayBiotech inc)

Journal: Journal of Personalized Medicine

Article Title: Human Papillomavirus Infection and EGFR Exon 20 Insertions in Sinonasal Inverted Papilloma and Squamous Cell Carcinoma

doi: 10.3390/jpm13040657

Figure Lengend Snippet: EGFR phosphorylation sites in IP-SCC. Eight phosphorylation sites in EGFR were investigated using an EGFR Phosphorylation Antibody Array 1 Assay. ( A ): Description of each well. ( B ): Results for IP-SCC1. ( C ): Results for IP-SCC-2. Of the eight sites, Y845 phosphorylation (F1 and F2 wells) was observed in both cases. Neg, negative control; Pos, positive control. Each antibody is spotted in duplicate vertically.

Article Snippet: A RayBio Human EGFR Phosphorylation Antibody Array 1 Assay (RayBiotech, Inc., Peachtree Corners, GA, USA) was used to screen specific EGFR phosphorylation sites (Y845, Y922, Y1045, Y1068, Y1086, Y1173, and Y1046/47) in two IP-SCC patients with ex20ins.

Techniques: Ab Array, Negative Control, Positive Control

Journal: International Journal of Biological Sciences

Article Title: Targeting the Sheddase Activity of ADAM17 by an Anti-ADAM17 Antibody D1(A12) Inhibits Head and Neck Squamous Cell Carcinoma Cell Proliferation and Motility via Blockage of Bradykinin Induced HERs Transactivation

doi: 10.7150/ijbs.9326

Figure Lengend Snippet: Effect of D1(A12) on the phosphorylation of HER family receptors and downstream signalling molecules under BK stimulation in HNSCC cells. (A) SCC13 cells were pre-incubated with different inhibitors for 2 h then treated with 10 nM BK for 10 min, after which the cell extracts were immunoblotted with the indicated antibodies. (B) SCC9 cells were treated in the same way as SCC13 cells and analysed with indicated antibodies. (C-D) Effect of ADAM17 siRNA and ADAM10 siRNA on the phosphorylation of EGFR and ERK in SCC13 cells. Cells were transfected with 3 different siRNAs and non-targeting siRNA. 72 h later, cells were serum starved for 48 h and then challenged with 10 nM BK for 10 min. After cell lysis, equal amount of proteins were analyzed by immunoblotting with indicated antibodies. (E-F) Effect of D1(A12) on the phosphorylation of HER family receptors in SCC13 cells. (E) Cells were treated as in A and analysed by immunoblotting with indicated antibodies. The same cell extract as 6E were exposed to the RayBio human EGFR phosphorylation antibody array 1 as described in M&M. (F) On each membrane, the left upper corner and right lower corner were spotted with positive control used for signal normalisation. The left lower corner was spotted with pan HER 1, 2 and 3. Five different phosphorylation sites of HER2, HER3 and HER4 were detected in the middle part of the membrane. (G) Normalisation of D1(A12) treated signals to human IgG treated signals. Signal intensity for a particular spot was divided by the average signal intensity of 4 positive controls on the same array membrane. Each phosphorylation site was spotted duplicate.

Article Snippet: To investigate whether BK can transactivate HER2, HER3 and HER4 along with EGFR and whether D1(A12) can prevent this induction in SCC13 cells, the RayBio human EGFR phosphorylation antibody array 1 was used to detect the relative levels of phosphorylation at specific sites of human HER family proteins.

Techniques: Incubation, Transfection, Lysis, Western Blot, Ab Array, Positive Control