Protein was extracted from cells in the 3D culture systems after 4 weeks of exposure to different stimuli, including 3D control, 3D+Hor, 3D+Mel, 3D+Hor+Mel, 3D+Hor+2-Iod, and 3D+Hor+Mel+Luz. ERK1/2 (ET160129, Huabio, China), p-ERK1/2 (ET160322, Huabio, China), DMC1 (ab11054, Abcam, UK), and β-actin (T56715S, Abmart, China) were examined by western blotting as described previously (Jia et al., 2013 (link)).
Et160129
Manufactured by Huabio
Sourced in China
The ET160129 is a laboratory equipment designed for conducting analytical and experimental procedures. It serves as a versatile tool for researchers and scientists in various fields. The core function of this product is to facilitate controlled and precise measurements during laboratory investigations.
Automatically generated - may contain errors
2 protocols using et160129
1
Protein Expression Analysis in 3D Cultures
2
Antibody-based Protein Interaction Assays
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Cell culture media DMEM and RPMI-1640, fetal bovine serum (FBS) were ordered from Gibco. Human recombinant EGF (Catalog number:10605-HNAE) was bought from Sino Biological company. ML792 (HY-108702), SUMOylation inhibitor 2-D08 (HY-114166), gefitinib (HY-50895), puromycin (HY-B1743A) and N-ethylmaleimide (NEM) (HY-D0843), were ordered from MedChemexpress company. Slurry anti-Flag M2 affinity gel (A2220) was ordered from Sigma. The protein-A beads (161–4013) were bought from Bio-Rad. The primary antibodies included individual monoclonal antibody of Myc (ab18185, Abcam), EGFR (sc-373746, Santa Cruz), AnxA6 (sc-166807, Santa Cruz). And rabbit monoclonal antibodies against Flag (BX00086), HA (BX00069-C3), His (BX00085-C3), SUMO1(ET1606-53), UBC9(ET1610-21), pY-EGFR(1086) (ET1612-30) and p-ERK1/2 (ET1610-13), ERK1/2 (ET1601-29), Cyclin D1(SA38-08) were all ordered from HuaBio Company in China. The antibody of β-tubulin (TA-10, Zsbio) and β-actin (TA-09, Zsbio) was used to quantify expression of housekeeping gene β-tubulin or β-actin for comparison normalization. The IgG antibody (A7016, Beyotime) was taken as nonspecific binding control for IP performance.
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