Human Bcl-2 Minus C-Terminus Antibody

Detection of Human Bcl‑2 Minus C-Terminus by Western Blot. Western blot shows lysates of KG-1 human acute myelogenous leukemia cell line. PVDF membrane was probed with 0.1-0.5 µg/mL of Mouse Anti-Human Bcl-2 Minus C-Terminus Monoclonal Antibody (Catalog # MAB827) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for Bcl-2 Minus C-Terminus at approximately 24 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of Human Bcl‑2 by Simple Western^TM. Simple Western lane view shows lysates of MCF‑7 human breast cancer cell line, loaded at 0.5 mg/mL. A specific band was detected for Bcl‑2 at approximately 24 kDa (as indicated) using 5 µg/mL of Mouse Anti-Human Bcl‑2 Minus C-Terminus Monoclonal Antibody (Catalog # MAB827). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Detection of Human Bcl-2 by Simple Western THZ1, suppresses Mcl-1 at the transcriptional level by disruption of its super-enhancer. (a) Real-time PCR analysis of Mcl1 mRNA levels in several GBM cells treated with vehicle or THZ1 for 24 h (n = 4). U87, U251, LN229, KNS42, and GBM22 cells were treated with 100 nM THZ1 while GBM14 was treated with 50 nM THZ1. Statistical significance was determined by two-tailed Student’s t-test; (b) U87 GBM cells were treated with DMSO or THZ1 100 nM for 24 h, subjected to CHIP with H3K27ac antibody and submitted for next generation sequencing. Super-enhancers were called using HOMER and depicted as a heatmap. The middle of each plot highlights the center of the super-enhancers (from −50 kb to 50 kb). The super enhancers are ranked by size and intensity levels are provided in the legend. The scale bar indicates the intensities. Blue depicts a high intensity level and red depicts a low intensity level; (c) A representation of global disruption of the super-enhancer landscape of U87 treated with DMSO or THZ1 100 nM in (b); (d) Shown are CHIP-seq (H3K27ac) tracks around the MCL1 locus (pile up values are indicated) in U87 treated with DMSO or THZ1 100 nM (super enhancer related to the Mcl-1 gene: chr1:150,601,879-150,630,909 (GRCh38/hg38)); (e) Standard western blots of cell lysates of U87 and GBM22 cells treated with increasing concentration of THZ1 for 24 h (pRpb1 corresponds to serine 5). Actin is used as a loading control. The protein expression levels were quantified using ImageJ (shown in cursive font); (f) Standard western blots or protein capillary electrophoresis of cell lysates of U87, U251, LN229, and GBM22 cells treated with increasing concentration of THZ1 for 24 h. Actin is used as a loading control in standard western blots and Vinculin is used as a loading control in protein capillary electrophoresis. The protein expression levels were quantified by using ImageJ (shown in cursive font). Uncropped blots are shown in Figure S10; (g) Shown are the protein expression levels of Mcl1, Noxa, Bcl2, and Bcl-xL following treatment with increasing concentration of THZ1 for 24 h in U87, U251, LN229, and GBM22 cells. FC: fold change. Shown are means and SD (n = 2–3). ***/**** p < 0.001. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/32752193), licensed under a CC-BY license. Not internally tested by R&D Systems.