primary anti bodies for gapdh  (Proteintech)

Journal: International Journal of Molecular Sciences

Article Title: Novel and Potent Small Molecules against Melanoma Harboring BRAF Class I/II/III Mutants for Overcoming Drug Resistance

doi: 10.3390/ijms22073783

Figure Lengend Snippet: The effect of SIJ1777 on AKT and MAPK signaling pathways in melanoma cell lines harboring various BRAF mutation status ( A ) SK-MEL-2 (wt) ( B ) SK-MEL-28 (class I) ( C ) C8161 (class II) ( D ) WM3670, WM3629 (class III). Cells were treated with 0.01, 0.1, 1 μM of SIJ1777, and 1 μM of vemurafenib, PLX8394, GNF-7, and SIJ1227 for 2 h. Cell lysates were subjected to western blot analysis to estimate the phospho- or total- form of AKT, MEK, ERK levels, and GAPDH was used as the internal loading controls.

Article Snippet: p-ERK1/2 (#5174), t-ERK1/2 (#9102), p-AKT(S473) (#9271), PARP (#9542), and GAPDH (#5174) primary anti-bodies were purchased from Cell signaling technology (Danvers, MA, USA). p-MEK1/2 antibody (sc-81503) was purchased from Santa Cruz (Dallas, TX, USA). t-AKT (A18120), t-MEK1/2 (A4868) antibodies were purchased from Abclonal.

Techniques: Protein-Protein interactions, Mutagenesis, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Novel and Potent Small Molecules against Melanoma Harboring BRAF Class I/II/III Mutants for Overcoming Drug Resistance

doi: 10.3390/ijms22073783

Figure Lengend Snippet: The effect of SIJ1777 on apoptosis induction. ( A ) Western blot for pro-apoptotic marker level (cleaved PARP) in melanoma cell lines. GAPDH was used as the internal loading control. ( B ) Quantification graphs of western blot results by ImageJ ( n = 3). ( C ) Apoptotic cell (annexin V-positive) population was measured by flow cytometry analysis against melanoma cell lines ( n = 3). Cells were treated with indicated substances for 24 h. Statistical significances were determined using a one-way ANOVA analysis (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001).

Article Snippet: p-ERK1/2 (#5174), t-ERK1/2 (#9102), p-AKT(S473) (#9271), PARP (#9542), and GAPDH (#5174) primary anti-bodies were purchased from Cell signaling technology (Danvers, MA, USA). p-MEK1/2 antibody (sc-81503) was purchased from Santa Cruz (Dallas, TX, USA). t-AKT (A18120), t-MEK1/2 (A4868) antibodies were purchased from Abclonal.

Techniques: Western Blot, Marker, Control, Flow Cytometry

Journal: International Journal of Molecular Sciences

Article Title: Novel and Potent Small Molecules against Melanoma Harboring BRAF Class I/II/III Mutants for Overcoming Drug Resistance

doi: 10.3390/ijms22073783

Figure Lengend Snippet: Clonogenic assay analysis of SIJ1777 in C8161. ( A , B ) 2D clonogenic assay (colony formation assay) results of the compounds on C8161 melanoma cell. After incubation with test compounds for 14 days, colonies were photographed without magnification. ( C , D ) 3D clonogenic assay (soft agar assay) results of test compounds on C8161 melanoma cell. Cells embedded within 0.35% low melting agar and incubated with the indicated compounds for 14 days and observed without magnification. ( B , D ) Number of colonies were determined automatically by ImageJ ( n = 3, respectively). ( E ) Western blot analysis of SIJ1777 in C8161. Cells were treated with 0.01, 0.1 μM of SIJ1777, and 0.1 μM of vemurafenib, PLX8394, GNF-7, and SIJ1227 for 24 h. Cell lysates were subjected to western blot analysis to estimate the phospho- or total- form of AKT, MEK, ERK levels, and GAPDH was used as the internal loading controls (left panel). Quantification result ( n = 3) of western blot result by ImageJ (right panel). Statistical significances were determined using a one-way ANOVA analysis (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001).

Article Snippet: p-ERK1/2 (#5174), t-ERK1/2 (#9102), p-AKT(S473) (#9271), PARP (#9542), and GAPDH (#5174) primary anti-bodies were purchased from Cell signaling technology (Danvers, MA, USA). p-MEK1/2 antibody (sc-81503) was purchased from Santa Cruz (Dallas, TX, USA). t-AKT (A18120), t-MEK1/2 (A4868) antibodies were purchased from Abclonal.

Techniques: Clonogenic Assay, Colony Assay, Incubation, Soft Agar Assay, Western Blot