mouse polyclonal antibodies against rhoa  (Proteintech)

Journal: Journal of Cellular and Molecular Medicine

Article Title: Use of iTRAQ ‐based quantitative proteomic identification of CHGA and UCHL1 correlated with lymph node metastasis in colorectal carcinoma

doi: 10.1111/jcmm.17793

Figure Lengend Snippet: The effect of chromogranin A (CHGA) and ubiquitin carboxyl‐terminal hydrolase isozyme L1 (UCHL1) silencing on Rho‐GTPase/AKT/NFκB pathway activation and the expression of EMT markers in CRC. After transfection with lentiviral shRNA targeting CHGA and UCHL1 (shCHGA and shUCHL1) or non‐targeting control (shControl), whole‐cell lysate proteins of HCT‐116 cells were subjected to western blotting, with antibodies against phosphorylation of Rho‐GTPase, AKT and NFκB p50 (A) as well as β‐catenin, cyclin E, twist 1/2 and vimentin (B) β‐Actin served as the loading control. The protein levels were quantified through densitometric analysis with the ratio of the untreated control (Control) set as onefold. The quantitative data are presented as the mean of three repeats from three independent experiments. * p < 0.05, compared with the control group.

Article Snippet: The antibodies were respectively obtained by Abcam Technology (Abcam) and Cell Signalling Technology, including mouse/rabbit polyclonal antibodies against RhoA Ser188, AKT Thr180Tyr182 and NFκB p50 bought from all culture materials were obtained from Gibco (Grand Island, NY, USA).

Techniques: Ubiquitin Proteomics, Activation Assay, Expressing, Transfection, shRNA, Control, Western Blot, Phospho-proteomics

Journal: Journal of Cellular and Molecular Medicine

Article Title: Use of iTRAQ ‐based quantitative proteomic identification of CHGA and UCHL1 correlated with lymph node metastasis in colorectal carcinoma

doi: 10.1111/jcmm.17793

Figure Lengend Snippet: Schematic presentation of the working model of CHGA and UCHL1 participate in promotion of the LNM‐associated CRC through endogenous Rho‐GTPase/AKT/NFκB signalling pathways‐mediated histone modification (H3K4me3) as reliable candidate LNM‐associated markers.

Article Snippet: The antibodies were respectively obtained by Abcam Technology (Abcam) and Cell Signalling Technology, including mouse/rabbit polyclonal antibodies against RhoA Ser188, AKT Thr180Tyr182 and NFκB p50 bought from all culture materials were obtained from Gibco (Grand Island, NY, USA).

Techniques: Modification

Journal: Journal of Cellular and Molecular Medicine

Article Title: Use of iTRAQ ‐based quantitative proteomic identification of CHGA and UCHL1 correlated with lymph node metastasis in colorectal carcinoma

doi: 10.1111/jcmm.17793

Figure Lengend Snippet: Alternation of histone modification of CHGA and UCHL1 promoters by the Rho‐GTPase/AKT/NFκB signalling pathways. After incubation with various concentrations of the specific inhibitors such as CCG‐1423, PDTC and wortmannin for 24 h, chromatin immunoprecipitation (ChIP) assays using antibodies against H3K4me3 was performed to pull down associated DNA in the HCT‐116 cells. Polymerase chain reaction amplified the precipitated DNA by using primer sets specific to the target sites (−1185 to −940 and −407 to −230) of CHGA and UCHL1 promoters. After normalized signal to the negative control ChIP asΔCT by subtracting the mean CT of the input from that of the individual region among the untreated control group, using the ΔΔCt method to calculate the effect of the specific inhibitors on specific genes. The data are presented as fold change of the untreated group (mean ± SD) of three independent experiments with * p < 0.01.

Article Snippet: The antibodies were respectively obtained by Abcam Technology (Abcam) and Cell Signalling Technology, including mouse/rabbit polyclonal antibodies against RhoA Ser188, AKT Thr180Tyr182 and NFκB p50 bought from all culture materials were obtained from Gibco (Grand Island, NY, USA).

Techniques: Modification, Incubation, Chromatin Immunoprecipitation, Polymerase Chain Reaction, Amplification, Negative Control, Control