Estrogen receptor alpha, Santa Cruz Biotechnology, sc-543

We generated a top 1000 list of the most studied and most published proteins and analyzed literature validation data for commercially available research antibodies. ESR1 encoding estrogen receptor alpha (ERα) is the ninth most studied gene on the list. In Sugiaman-Trapman et al., BMC Genomics (2018), the authors perform ESR1 siRNA KD in MCF7 cells with a scrambled siRNA control. Antibody sc-543 (1:500) is used for WB using whole-cell lysates (40–80 µg), collected 24 h after transfection. The data are shown in Figure 5c. The antibody detects a band that is substantially reduced after KD supporting specific detection of ERα by sc-543 in WB under the tested conditions. The blot is cropped and lacks molecular-weight markers, limiting assessment of band size and nonspecific bands. We highly encourage other scientists to share their observations.

We generated a top 1000 list of the most studied and most published proteins and analyzed literature validation data for commercially available research antibodies. ESR1 encoding estrogen receptor alpha (ERα) is the ninth most studied gene on the list. In Glont et al., PLOS ONE (2019), the authors compare sc-543 head-to-head with two other ERα antibodies using ChIP-seq and RIME (IP–MS). Both assays use nuclear/chromatin preparations from crosslinked MCF7 cells, with IgG controls. ChIP-seq (10 µg antibody) shows similar ERα binding profiles across antibodies. However, sc-543 detects 124 peaks in ERα-negative MDA-MB-231 cells, indicating higher background (Figure 1). RIME confirms capture of ERα and known associated proteins, with similar performance across antibodies (Figure 2). These data support specific enrichment of ERα in ChIP and RIME (IP–MS) under the tested conditions, although sc-543 shows more background in ChIP. No KO/KD controls are included. The authors report that sc-543 has been discontinued. We highly encourage other scientists to share their observations.