POU domain class 2-associating factor 1 is a protein that in humans is encoded by the POU2AF1 gene. Transcriptional coactivator that specifically associates with either OCT1 or OCT2. It boosts the OCT1 mediated promoter activity and to a lesser extent, that of OCT2. It has no intrinsic DNA-binding activity. It recognizes the POU domains of OCT1 and OCT2. It is essential for the response of B-cells to antigens and required for the formation of germinal centers. Reduced activation of Bob1-deficient B cells following antigenic stimulation is often reported. The action of Bob1 on B cell maturation and tolerance could be mediated by effects on B cell activation, in part by regulating the expression of immunosuppressive miRNAs. Bob1 has long been considered a B cell‐specific factor that interacts with the transcription factors Oct1 and Oct2 to enhance octamer‐dependent transcription. Bob1 can directly bind to and transactivate the promoters of Bcl6 and Btla. The expression of BOB1 has proven useful for identifying certain lymphomas as being B-cell lymphomas, as exemplified in studies which use BAB1 expression to help identify lymphomas as being diffuse large B-cell lymphomas, not otherwise specified.
Background References
1. Galiegue-Zouitina S et al. Fusion of the LAZ3/BCL6 and BOB1/OBF1 genes by t(3; 11) (q27; q23) chromosomal translocation. C R Acad Sci III Sci Vie 318:1125-1131 (1995).
2. John M. Lindner et al. A C-Terminal Acidic Domain Regulates Degradation of the Transcriptional Coactivator Bob1. Mol Cell Biol 33(23):4628-4640 (2013).
Sequence Similarity
Belongs to the POU2AF1 family.
Tissue Specificity
B-cell specific.
Post-translational Modification
Ubiquitinated; mediated by SIAH1 or SIAH2 and leading to its subsequent proteasomal degradation.
Western blot analysis of BOB1 on different lysates with Rabbit anti-BOB1 antibody (ET7109-68) at 1/5,000 dilution.
Lane 1: Daudi (Human Burkitt's lymphoma cell) cell lysate Lane 2: Raji (Human Burkitt's lymphoma cell) cell lysate Lane 3: Ramos (Human Burkitt's lymphoma cell) cell lysate
Lysates/proteins at 20 µg/Lane. Exposure time: 4 seconds; ECL: K1801
Blocking: 5% NFDM/TBST, 1 hour at room temperature Primary antibody: ET7109-68, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃ Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature
Predicted band size: 27 kDa Observed band size: 36 kDa
Immunohistochemical analysis of paraffin-embedded human B-cell lymphoma tissue with Rabbit anti-BOB1 antibody (ET7109-68) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7109-68) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-BOB1 antibody (ET7109-68) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7109-68) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Flow cytometric analysis of BOB1 was done on THP-1 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET7109-68, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
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