Cell Signaling Technology
XP Monoclonal Antibody

Product Pathways - Lymphocyte Signaling

Bcl-11B (D6F1) XP® Rabbit mAb #12120

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP IF-IC ChIP H (M) (R) (Mk) Endogenous 120-130 Rabbit IgG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IF-IC=Immunofluorescence (Immunocytochemistry)  ChIP=Chromatin IP
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

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

Source / Purification

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

Western Blotting

Western Blotting

Western blot analysis of extracts from MOLT-4 and Jurkat cells using Bcl-11B (D6F1) XP® Rabbit mAb.

IP

IP

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.

IF-IC

IF-IC

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 IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 Jurkat cells and either 10 μl of Bcl-11B (D6F1) XP® Rabbit mAb or 2 μl of 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.

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.

Application References

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For Research Use Only. Not For Use In Diagnostic Procedures.

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