Human/Mouse EBF-1 Antibody

Detection of Human/Mouse EBF‑1 by Western Blot. Western blot shows lysates of Raji human Burkitt's lymphoma cell line, Daudi human Burkitt's lymphoma cell line, and CH-1 mouse B cell lymphoma cell line, and L1.2 mouse pro-B cell line. PVDF membrane was probed with 2 µg/mL of Human/Mouse EBF-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5165) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for EBF-1 at approximately 70 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of Human EBF-1 by Western Blot Differential binding of transcription factors on EBV genomes in type I and type III latency.(A) ChIP-Seq tracks mapped to EBV genome in LCL (red) or Mutu I (blue) for EBF1 or RBP-j kappa as indicated. EBNA2 ChIP-Seq for LCL (black). Y-axis represent raw read counts. (B) ChIP-qPCR for EBF1 or RBP-j kappa in LCL (red) or Mutu I (blue) at various EBV genome regulatory regions as indicated and Actin genomic region as negative control. (C) Same as in B, except for isogenic EBV positive BL cells Kem III (red) or Kem I (blue). Asterisk indicates p < 0.05. (D) Western blot of Mutu I, LCL, Kem I, and Kem III probed with antibody to EBF1, RBP-j kappa, LMP1, and EBNA2, with cellular loading controls for GAPDH and H3K4me3, as indicated. (E) RT-qPCR for EBV type III gene expression in LCL or Mutu I (left panel) or Kem I and Kem III (right panel). Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/26752713), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human EBF-1 by Western Blot Cooperative binding and colocalization of EBF1, RBP-j kappa, and EBNA2 at DNA regulatory elements.(A) DNA-affinity binding assays with nuclear extracts from EREB2.5 cells with (+) or without (-) estradiol (E2) using DNA regulatory elements from IL7, HES1 (left), or LMP1, Qp (middle), or TOM1 (right). Input (10%) is indicated, and bound proteins were assayed by Western blot for EBF1, RBP-j kappa, EBNA2, and specificity controls for PARP1 and Actin. (B) ChIP-reChIP assays in LCLs with EBF1 as first ChIP, followed by a reChIP with either no antibody control or antibody to EBF1, RBP-j kappa or EBNA2. ReChIP DNA was quantified by qPCR at LMP1, IL7, and BDH2 (an EBF1 only site) binding sites. (C) Same as in B, except first ChIP was with RBP-j kappa followed by a ReChIP with no antibody control, or EBF1, RBP-j kappa, or EBNA2 antibody and assayed at LMP1, IL7, or LZTFL1 (an RBP-j kappa only site) binding sites. Asterisk indicates p < 0.05. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/26752713), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human EBF-1 by Western Blot EBNA2-dependent transcription factor redistribution in chromatin occupancy.EREB2.5 cells were treated with (+) or without (-) estradiol (E2) for 24 or 48 hrs and then assayed by ChIP for binding to EBF1 (A and B), RBP-j kappa (C and D), or EBNA2 (E and F) at cellular (A, C, E) or EBV genome sites (B, D, F). Actin genomic region (cellular) or Qp (EBV) was used as negative binding control for EBF1, RBP-j kappa, ορ EBNA2 ChIP. (G) ChIP binding for CTCF, PU.1, or PAX5 in EREB2.5 cells treated (blue) or untreated (red) with E2 for 48 hrs. PPP1R1B or KCTD17 genomic region was negative binding control PU.1 or PAX5, respectively. Asterisk indicates p < 0.05. (H) Western blot showing protein levels for EBF1, RBP-j kappa, EBNA2, Actin, and CTCF in EREB2.5 cells at 24 and 48 hrs after E2 withdrawal. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/26752713), licensed under a CC-BY license. Not internally tested by R&D Systems.