Cell Signaling Technology

Product Pathways - Transcription Factors

NKX3.1 (D2Y1A) XP® Rabbit mAb #83700

CaP  

No. Size Price
83700S 100 µl ( 10 western blots ) ¥3,580.00 现货查询 购买询价
83700T 20 µl ( 2 western blots ) ¥1,400.00 现货查询 购买询价
83700 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human, Endogenous 30 Rabbit IgG
IF-IC 1:100
ChIP 1:100

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IF-IC=Immunofluorescence (Immunocytochemistry), ChIP=Chromatin IP,

Specificity / Sensitivity

NKX3.1 (D2Y1A) XP® Rabbit mAb recognizes endogenous levels of total NKX3.1 protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein specific to human NKX3.1 protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from human prostate and various cell lines using NKX3.1 (D2Y1A) XP® Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). DND-41 cell are negative for NKX3.1 expression, confirming specificity of the antibody.

IF-IC

IF-IC

Confocal immunofluorescent analysis of LNCaP (left; positive) and HeLa (right; negative) cells using NKX3.1 (D2Y1A) XP® Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 LNCaP cells and either 5 μl of NKX3.1 (D2Y1A) 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 CYP2B7P Pseudogene Promoter Primers #84846, human MALSU1 downstream 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

NKX3.1 is a homeobox transcription factor that in mammals plays a defining role in embryonic prostate morphogenesis. The expression of mammalian NKX3.1 is androgen-dependent, restricted primarily to developing and mature prostate epithelium, and is frequently reduced or lost in prostate cancer (1-3). The human NKX3.1 gene is located on chromsome 8p21.2, within a region that shows loss of heterozygosity (LOH) in >50% of prostate cancer cases (2). Allelic loss at the NKX3.1 locus is also common in high grade Prostate Intraepithelial Neoplasia (PIN), thought to be a putative precursor lesion to invasive prostate adenocarcinomas, suggesting that LOH at the NKX3.1 locus is a critical early step in prostate cancer development (4). Notably, the remaining NKX3.1 allele is intact in the majority of LOH cases, leading to the suggestion that NKX3.1 functions as a haploinsufficient tumor suppressor (4-6). Due to its highly restricted expression in prostate epithelial cells, NKX3.1 has been suggested as a diagnostic marker of prostate carcinoma (7), and may have additional utility as a biomarker of metastatic lesions originating in the prostate (8).

  1. Bhatia-Gaur, R. et al. (1999) Genes Dev 13, 966-77.
  2. He, W.W. et al. (1997) Genomics 43, 69-77.
  3. Bowen, C. et al. (2000) Cancer Res 60, 6111-5.
  4. Magee, J.A. et al. (2003) Cancer Cell 3, 273-83.
  5. Voeller, H.J. et al. (1997) Cancer Res 57, 4455-9.
  6. Bethel, C.R. et al. (2006) Cancer Res 66, 10683-90.
  7. Epstein, J.I. et al. (2014) Am J Surg Pathol 38, e6-e19.
  8. Conner, J.R. and Hornick, J.L. (2015) Adv Anat Pathol 22, 149-67.

Application References

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Protocols

Companion Products


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

DRAQ5 is a registered trademark of Biostatus Limited.

DyLight is a trademark of Thermo Fisher Scientific, Inc. and its subsidiaries.

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

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

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

Tween is a registered trademark of ICI Americas, Inc.

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

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