Revision 5

#12120

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CST Logo
Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

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

Applications:
W, IP, IF-F, IF-IC, ChIP, ChIP-seq

Reactivity:
H M

Sensitivity:
Endogenous

MW (kDa):
120-130

Source/Isotype:
Rabbit IgG

UniProt ID:
#Q9C0K0

Entrez-Gene Id:
64919

Product Usage Information

For optimal ChIP and ChIP-seq results, use 10 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.

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:100
Immunofluorescence (Frozen) 1:100
Immunofluorescence (Immunocytochemistry) 1:200
Chromatin IP 1:50
Chromatin IP-seq 1:50

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 #16148.

Specificity/Sensitivity

Bcl-11B (D6F1) XP® Rabbit mAb recognizes endogenous levels of total Bcl-11B protein.

Species predicted to react based on 100% sequence homology

Rat, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu391 of human Bcl-11B protein.

Background

Bcl-11B (Ctip2) is a COUP-TF interacting protein that belongs to the C2H2 type zinc finger protein family (1). Bcl-11B is highly expressed in the brain and is critical for the development of neurons, as well as other tissues and organs. Bcl-11B also plays an essential role in T cell lineage commitment and maintenance of T cell identity (1-3). Two isoforms of Bcl-11B are found to be encoded by the BCL11B gene, possibly through exon-skipping (4). Bcl-11B is a transcription factor which binds to target genes through the 2nd and 3rd zinc-finger domains of exon 4 (3), while also interacting with various protein partners including COUP-TF proteins (1), the NuRD complex (5,6), HDAC1, HDAC2, and SUV39H1 (7). Research studies have shown that mutations and deletion of Bcl-11B contribute to the development of thymic lymphoma in mice and T cell acute lymphoblastic leukemia in humans, indicating a role as a tumor suppressor (4,8). Mechanistic studies have shown that Bcl-11B represses gene expression of the E3 ubiquitin ligase HDM2 in a p53-dependent manner (9).

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 IF-F: Immunofluorescence (Frozen) IF-IC: Immunofluorescence (Immunocytochemistry) ChIP: Chromatin IP ChIP-seq: Chromatin IP-seq

Cross-Reactivity Key

H: Human M: Mouse

Trademarks and Patents

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

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

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 5

Cell Signaling Technology Logo
Western blot analysis of extracts from MOLT-4 and Jurkat cells using Bcl-11B (D6F1) XP® Rabbit mAb.
Western Blotting Image 1: Bcl-11B (D6F1) XP<sup>®</sup> Rabbit mAb
Immunoprecipitation of Bcl-11B from MOLT-4 cell extracts, using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or Bcl-11B (D6F1) XP® Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using Bcl-11B (D6F1) XP® Rabbit mAb.
Immunoprecipitation Image 1: Bcl-11B (D6F1) XP<sup>®</sup> Rabbit mAb
Confocal immunofluorescent analysis of fixed frozen postnatal day 5 (P5) mouse dentate gyrus (left), striatum (middle), and frontal cortex (right) using Bcl-11B (D6F1) XP® Rabbit mAb #12120 (green) and ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Immunofluorescence Image 1: Bcl-11B (D6F1) 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 5

Cell Signaling Technology Logo
Confocal immunofluorescent analysis of fixed frozen adult mouse dentate gyrus (left), striatum (middle), and frontal cortex (right) using Bcl-11B (D6F1) XP® Rabbit mAb #12120 (green) and ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Immunofluorescence Image 2: Bcl-11B (D6F1) XP<sup>®</sup> Rabbit mAb
Confocal immunofluorescent analysis of Jurkat (positive; left) and HeLa (negative; right) cells using Bcl-11B (D6F1) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Immunofluorescence Image 1: Bcl-11B (D6F1) XP<sup>®</sup> Rabbit mAb
Chromatin immunoprecipitations were performed with cross-linked chromatin from Jurkat cells and Bcl-11B (D6F1) 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 IL9 gene. For additional ChIP-seq tracks, please download the product data sheet.
Chromatin Immunoprecipitation Image 1: Bcl-11B (D6F1) 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 5

Cell Signaling Technology Logo
Chromatin immunoprecipitations were performed with cross-linked chromatin from Jurkat cells and Bcl-11B (D6F1) 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 1 (upper), including IL9 gene (lower).
Chromatin Immunoprecipitation Image 2: Bcl-11B (D6F1) XP<sup>®</sup> Rabbit mAb
Chromatin immunoprecipitations were performed with cross-linked chromatin from Jurkat cells and eitherBcl-11B (D6F1) 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 CDKN1B Promoter Primers #11951, human MDM2 promoter primers, 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: Bcl-11B (D6F1) 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.