Revision 5

#12120Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, 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 Information

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 Reactivity:

Human, Mouse

Species predicted to react based on 100% sequence homology

Mouse, 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).

  1. Avram, D. et al. (2000) J Biol Chem 275, 10315-22.
  2. Liu, P. et al. (2010) Immunol Rev 238, 138-49.
  3. Kominami, R. (2012) Proc Jpn Acad Ser B Phys Biol Sci 88, 72-87.
  4. Wakabayashi, Y. et al. (2003) Biochem Biophys Res Commun 301, 598-603.
  5. Cismasiu, V.B. et al. (2005) Oncogene 24, 6753-64.
  6. Topark-Ngarm, A. et al. (2006) J Biol Chem 281, 32272-83.
  7. Marban, C. et al. (2007) EMBO J 26, 412-23.
  8. Gutierrez, A. et al. (2011) Blood 118, 4169-73.
  9. Obata, M. et al. (2012) Cell Signal 24, 1047-52.

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

WB: 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 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 All: all species expected

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.

使用に関する制限

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Revision 5
#12120

Bcl-11B (D6F1) XP® Rabbit mAb

Western Blotting Image 1: Bcl-11B (D6F1) XP® Rabbit mAb Expand Image
Western blot analysis of extracts from MOLT-4 and Jurkat cells using Bcl-11B (D6F1) XP® Rabbit mAb.
Immunoprecipitation Image 1: Bcl-11B (D6F1) XP® Rabbit mAb Expand Image
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.
Immunofluorescence Image 1: Bcl-11B (D6F1) XP® Rabbit mAb Expand Image
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 2: Bcl-11B (D6F1) XP® Rabbit mAb Expand Image
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 1: Bcl-11B (D6F1) XP® Rabbit mAb Expand Image
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).
Chromatin Immunoprecipitation Image 1: Bcl-11B (D6F1) XP® Rabbit mAb Expand Image
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 2: Bcl-11B (D6F1) XP® Rabbit mAb Expand Image
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 3: Bcl-11B (D6F1) XP® Rabbit mAb Expand Image
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.