Revision 5
#5650
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For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:
W, IP, ChIP, C&R
Reactivity:
H M
Sensitivity:
Endogenous
MW (kDa):
87-98
Source/Isotype:
Rabbit IgG
UniProt ID:
#P41182
Entrez-Gene Id:
604
Product Usage Information
The CUT&RUN dilution was determined using CUT&RUN Assay Kit #86652.
Application | Dilution |
---|---|
Western Blotting | 1:1000 |
Immunoprecipitation | 1:200 |
Chromatin IP | 1:200 |
CUT&RUN | 1:25 |
Storage
Specificity/Sensitivity
BCL6 (D65C10) Rabbit mAb recognizes endogenous levels of total BCL6 protein.
Source / Purification
Monoclonal antibody is produced by immunizing animals with a recombinant protein specific to human BCL6 protein.
Background
Chromosomal translocations result in misregulation of the proto-oncogene BCL6 in patients with B cell-derived non-Hodgkin's lymphoma (1). The BCL6 gene is selectively expressed in mature B cells and encodes a nuclear phosphoprotein that belongs to the BTB/POZ zinc finger family of transcription factors (2,3). BCL6 protein can bind to target DNA sequences of Stat6 and, analogous to Stat6, modulate the expression of interleukin-4-induced genes (4). Furthermore, BCL6 restrains p53-dependent senescence, making BCL6-active tumors functionally p53-negative (5). The mitogen-activated protein kinases, Erk1 and Erk2, but not JNK, phosphorylate BCL6 at multiple sites. Phosphorylation of BCL6 at Ser333 and Ser343 results in degradation of BCL6 by the ubiquitin/proteasome pathway in B cells (6,7). In addition, BCL6 is acetylated and its transcriptional repressor function is inhibited by the transcriptional co-activator p300 (8).
Background References
- Ye, B.H. et al. (1993) Science 262, 747-50.
- Onizuka, T. et al. (1995) Blood 86, 28-37.
- Zollman, S. et al. (1994) Proc Natl Acad Sci U S A 91, 10717-21.
- Harris, M.B. et al. (1999) Mol Cell Biol 19, 7264-75.
- Shvarts, A. et al. (2002) Genes Dev 16, 681-6.
- Moriyama, M. et al. (1997) Oncogene 14, 2465-74.
- Niu, H. et al. (1998) Genes Dev 12, 1953-61.
- Bereshchenko, O.R. et al. (2002) Nat Genet 32, 606-13.
Species Reactivity
Species reactivity is determined by testing in at least one approved application (e.g., western blot).
Western Blot Buffer
IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
Applications Key
W: Western Blotting IP: Immunoprecipitation ChIP: Chromatin IP C&R: CUT&RUN
Cross-Reactivity Key
H: Human M: Mouse R: Rat Hm: Hamster Mk: Monkey Vir: Virus Mi: Mink C: Chicken Dm: D. melanogaster X: Xenopus Z: Zebrafish B: Bovine Dg: Dog Pg: Pig Sc: S. cerevisiae Ce: C. elegans Hr: Horse GP: Guinea Pig Rab: Rabbit G: Goat All: All Species Expected
Trademarks and Patents
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.
Limited Uses
Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.
Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.
Revision 5
#5650
BCL6 (D65C10) Rabbit mAb
Western blot analysis of extracts from various cell lines using BCL6 (D65C10) Rabbit mAb.
Chromatin immunoprecipitations were performed with cross-linked chromatin from Raji cells and either BCL6 (D65C10) Rabbit mAb, or Normal Rabbit IgG #2729, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human BCL2 Promoter Primers #12924, SimpleChIP® Human HSP90B1 Promoter Primers #12899, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
CUT&RUN was performed with Raji cells and BCL6 (D65C10) Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA library was prepared using DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #56795. The figure shows binding across USP5 gene, a known target gene of BCL6 (see additional figure containing CUT&RUN-qPCR data).
Revision 5
#5650
BCL6 (D65C10) Rabbit mAb
CUT&RUN was performed with Raji cells and BCL6 (D65C10) Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA library was prepared using DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #56795. The figures show binding across chromosome 12 (upper), including USP5 (lower), a known target gene of BCL6 (see additional figure containing CUT&RUN-qPCR data).
CUT&RUN was performed with Raji cells and either BCL6 (D65C10) Rabbit mAb or Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362, using CUT&RUN Assay Kit #86652. The enriched DNA was quantified by real-time PCR using human SIN3A promoter primers, human USP5 promoter primers, and human MYOM3 promoter primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.