Human BACH1 Antibody

Detection of Human BACH1 by Western Blot.

Western blot shows lysates of K562 human chronic myelogenous leukemia cell line and A172 human glioblastoma cell line. PVDF membrane was probed with 0.2 µg/mL of Human/Mouse BACH1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5776) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for BACH1 at approximately 110 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

BACH1 in Human Lung Cancer Tissue.

BACH1 was detected in immersion fixed paraffin-embedded sections of human lung cancer tissue using Human/Mouse BACH1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5776) at 10 µg/mL overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Detection of BACH1 by Immunohistochemistry

BACH1 affects the sensitivity of ESCC cells to ferroptosis.A Relative expression of the BACH1, SLC7A11, GPX4, and ACSL4 proteins in esophageal epithelium Het-1A and ESCC (KYSE150, KYSE170, KYSE180, KYSE30, KYSE410 and KYSE510) cells as determined by western blotting. beta -actin was used as a loading control. B Concentration–response plot of different ESCC cells to RSL3, a ferroptosis-triggering agent, as determined by CCK-8 assay with measurement of the absorbance at 450 nm. C-F Correlations between BACH1 (C), SLC7A11 (D), GPX4 (E), and ACSL4 (F) protein expression levels (A) and their relevant IC50 values of RSL3 (B) against ESCC cells. G Lentivirus-mediated stable ectopic expression (BACH1-OE and shBACH1) and knockdown (shCtrl and shBACH1) of BACH1 in KYSE150 and KYSE170 cells. The efficiency of overexpression and knockdown was determined by western blotting. Long and short exposures (exp.) of the BACH1 blots are shown for KYSE150 cells. H Forced expression of BACH1 decreased the IC50 value of RSL3 against KYSE150 cells (left panel), whereas knockdown BACH1 knockdown increased the IC50 value of RSL3 against KYSE170 cells (right panel). I Flow cytometric analysis after C11-BODIPY591/581 staining showed the changes in lipid peroxidation after transient perturbation of BACH1 followed by treatment with 1 µM RSL3 in KYSE150 cells. J Phase-contrast microscopic changes in cellular shape in the absence (DMSO) or presence of 1 µM RSL3 for 6 h. K Ultrastructural changes in vector control and BACH1-OE cells as determined through transmission electron microscopy after incubation in the absence or presence of 1 µM RSL3 for 6 h. The red arrows indicate mitochondria. Scale bar, 0.5 μm. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36670112), licensed under a CC-BY license. Not internally tested by R&D Systems.