Revision 7

#14895

Store at -20C

BCL6 (D4I2V) XP® Rabbit mAb
CST Logo
Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

Web:

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Applications:
W, W-S, IP, IF-IC, FC-FP, ChIP, ChIP-seq, C&R

Reactivity:
H

Sensitivity:
Endogenous

MW (kDa):
87-98

Source/Isotype:
Rabbit IgG

UniProt ID:
#P41182

Entrez-Gene Id:
604

Product Usage Information

For optimal ChIP and ChIP-seq results, use 5 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.

The CUT&RUN dilution was determined using CUT&RUN Assay Kit #86652.

Application Dilution
Western Blotting 1:1000
Simple Western™ 1:10 - 1:50
Immunoprecipitation 1:200
Immunofluorescence (Immunocytochemistry) 1:50 - 1:100
Flow Cytometry (Fixed/Permeabilized) 1:800 - 1:1600
Chromatin IP 1:100
Chromatin IP-seq 1:100
CUT&RUN 1:100

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

For a carrier free (BSA and azide free) version of this product see product #47899.

Specificity/Sensitivity

BCL6 (D4I2V) XP® Rabbit mAb recognizes endogenous levels of total BCL6 protein. It is predicted to detect both human BCL6 isoform 1 and isoform 2.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a recombinant protein fragment spanning residues Asp210 through Thr409 of 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

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 nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

W: Western Blotting W-S: Simple Western™ IP: Immunoprecipitation IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized) ChIP: Chromatin IP ChIP-seq: Chromatin IP-seq C&R: CUT&RUN

Cross-Reactivity Key

H: Human

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

Alexa Fluor is a registered trademark of Life Technologies Corporation.

All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

限制使用

除非 CST 的合法授书代表以书面形式书行明确同意，否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件，除非书 CST 的合法授书代表以书面形式书独接受， 否书均被拒书，并且无效。

专品专有“专供研究使用”的专专或专似的专专声明， 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的， 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专，且专用于研专用途。将专品用于专断、专防或治专目的， 或专专售（专独或作专专成）或其他商专目的而专专专品，均需要 CST 的专独专可。客专：(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品，或使用专品制造任何商专专品，(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专，或使用专品开专任何与 CST 的专品或服专专争的专品或服专， (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专，(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品 ， (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专

Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 7

#14895

BCL6 (D4I2V) XP® Rabbit mAb

Cell Signaling Technology Logo
Western blot analysis of extracts from various cell lines using BCL6 (D4I2V) XP® Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western Blotting Image 1: BCL6 (D4I2V) XP<sup>®</sup> Rabbit mAb
Simple Western™ analysis of lysates (0.1 mg/mL) from Daudi cells using BCL6 (D4I2V) XP® Rabbit mAb #14895. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ ​​​​​​​ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
Western Blotting Image 1: BCL6 (D4I2V) XP<sup>®</sup> Rabbit mAb
Immunoprecipitation of BCL6 from Ramos cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or BCL6 (D4I2V) XP® Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using BCL6 (D4I2V) XP® Rabbit mAb. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb #3678 was used as a secondary antibody.
Immunoprecipitation Image 1: BCL6 (D4I2V) XP<sup>®</sup> Rabbit mAb
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 7

#14895

BCL6 (D4I2V) XP® Rabbit mAb

Cell Signaling Technology Logo
Confocal immunofluorescent analysis of Ramos (left) and Jurkat (right) cells using BCL6 (D4I2V) XP® Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red).
Immunofluorescence Image 1: BCL6 (D4I2V) XP<sup>®</sup> Rabbit mAb
Flow cytometric analysis of Jurkat cells (blue) and Ramos cells (green) using BCL6 (D4I2V) XP® Rabbit mAb (solid lines) or a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
Flow Cytometry Image 1: BCL6 (D4I2V) XP<sup>®</sup> Rabbit mAb
Chromatin immunoprecipitations were performed with cross-linked chromatin from Raji cells and BCL6 (D4I2V) XP® Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across HSP90AB1/HSP90β, a known target gene of BCL6 (see additional figure containing ChIP-qPCR data).
Chromatin Immunoprecipitation Image 1: BCL6 (D4I2V) XP<sup>®</sup> Rabbit mAb
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 7

#14895

BCL6 (D4I2V) XP® Rabbit mAb

Cell Signaling Technology Logo
Chromatin immunoprecipitations were performed with cross-linked chromatin from Raji cells and BCL6 (D4I2V) XP® Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across chromosome 6 (upper), including HSP90AB1/HSP90β (lower), a known target gene of BCL6 (see additional figure containing ChIP-qPCR data).
Chromatin Immunoprecipitation Image 2: BCL6 (D4I2V) XP<sup>®</sup> Rabbit mAb
Chromatin immunoprecipitations were performed with cross-linked chromatin from Raji cells and either BCL6 (D4I2V) XP® 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 HSP90β 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.
Chromatin Immunoprecipitation Image 3: BCL6 (D4I2V) XP<sup>®</sup> Rabbit mAb
CUT&RUN was performed with Raji cells and BCL6 (D4I2V) XP® 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 GSK3A gene, a known target gene of BCL6 (see additional figure containing CUT&RUN-qPCR data).
CUT & RUN Image 1: BCL6 (D4I2V) XP<sup>®</sup> Rabbit mAb
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 7

#14895

BCL6 (D4I2V) XP® Rabbit mAb

Cell Signaling Technology Logo
CUT&RUN was performed with Raji cells and BCL6 (D4I2V) XP® 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 19 (upper), including GSK3A (lower), a known target gene of BCL6 (see additional figure containing CUT&RUN-qPCR data).
CUT & RUN Image 2: BCL6 (D4I2V) XP<sup>®</sup> Rabbit mAb
CUT&RUN was performed with Raji cells and either BCL6 (D4I2V) XP® 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 GSK3A promoter primers, human GNB2 promoter primers, and human OCM2 exon 1 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.
CUT & RUN Image 3: BCL6 (D4I2V) XP<sup>®</sup> Rabbit mAb
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.