hfe hemochromatosis polyclonal antibody  (Bioss)

Journal: Oncotarget

Article Title: Tumor-initiating cells of breast and prostate origin show alterations in the expression of genes related to iron metabolism

doi: 10.18632/oncotarget.14093

Figure Lengend Snippet: Expression of the HEPH g ene at the mRNA level in breast non-malignant cell line MCF10A, in TICs derived from breast cancer cell lines MCF-7, BT-474, T-47D and ZR-75-30 as well as from prostate cancer cell lines DU-145 and LNCaP has been determined ( A ) together with protein levels in the MCF-7 cell line (CTRL) and MCF-7 derived spheres (SPH) ( B ). Similarly, the expression of the HFE gene at the mRNA ( C ) level as well as protein level ( D ) in TICs is documented. Experiments were performed at least in triplicate, standard error is SEM, p -values lower than 0.05 are denoted with a star and were calculated by the GenEx software using the unpaired t-test and plotted with GraphPad prism software. Number sign denotes statistical significance involving Dun-Bonferroni correction. The protein expression was quantified by the image J software from 2 to 5 independent samples, standard error is SEM, p -values lower than 0.05 are denoted with a star and were calculated and plotted in GraphPad prism, using the unpaired t-test .

Article Snippet: Gels were then separated according to standard procedure at 20 mA per well, washed in 1× towbin buffer and blotted onto a nitrocellulose membrane via Xcell blotting module (Invitrogen) at constant voltage (35 V) for 2 h. Membranes were then blocked with 5% non-fat milk for 1 h, washed and incubated in 5% bovine serum albumin (BSA)/Tris-buffered saline (TBS)/0,1% Tween-20 with primary antibodies against ABCB10 (ThermoScientific #PA5-30468, dil. 1:1000), ACO1 (ThermoScientific #PA5-27824, dil. 1:1000), CYBRD1 (Bioss #bs-8297R, dil. 1:1000), EPAS1 (ThermoScientific #PA116510, dil. 1:1000), GLRX5 (Bioss #bs-13395R, dil. 1:1000), HEPH (Bioss #bs-15458R, dil. 1:1000), HFE (Bioss #bs-12335R, dil. 1:1000), IREB2 (ThermoScientific #PA116544, 1:500), QSOX1 (Sigma #SAB2700031, dil. 1:1000), TFRC (ThermoScientific #13-6800, dil. 1:2500), SLC39A14 (Abcam #ab191199, dil. 1:2000), SLC40A1 (Bioss #bs-4906R, dil. 1:1000) SLC11A2 (Cell Signalling #15083, dil. 1:1000), Ferritin (Abcam #ab75973, dil. 1:1000), Actin (Thermoscientific #MA5-15739-HRP, dil. 1:2000), Tubulin (Abcam #ab4742, dil. 1:5000) overnight.

Techniques: Expressing, Derivative Assay, Software