Cell Signaling Technology

Product Pathways - Nuclear Receptor Signaling

Vitamin D3 Receptor (D2K6W) Rabbit mAb #12550

Nuclear Hormone Receptors   sc-1008   sc-13133   Steroid Receptors   VDR   Vitamin D Receptor   Vitamin D3 Receptor  

No. Size Price
12550S 100 µl ( 10 western blots ) ¥3,100.00 现货查询 购买询价
12550 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse, Endogenous 48, 54 Rabbit IgG
IP 1:100
IHC-P 1:200
ChIP 1:50

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, IHC-P=Immunohistochemistry (Paraffin), ChIP=Chromatin IP,

Homology

Species predicted to react based on 100% sequence homology: Hamster, Bovine, Pig, Horse,

Specificity / Sensitivity

Vitamin D3 Receptor (D2K6W) Rabbit mAb recognizes endogenous levels of total vitamin D3 receptor protein. This antibody does not cross-react with vitamin D3 receptor-like proteins. Based upon sequence alignment, this antibody is predicted to react with both VDRB1 and VDRB2 isoforms.Vitamin D3 Receptor (D2K6W) Rabbit mAb能够识别内源性水平的总维生素D3受体蛋白。该抗体不与维生素D3受体样蛋白发生交叉反应。基于序列对比,预测该抗体与VDRB1 和VDRB2亚型发生反应。

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of human vitamin D receptor isoform A protein.该单克隆抗体通过用合成肽免疫动物制备,该合成肽是人维生素D3受体蛋白A亚型靠近氨基末端附近的残基。

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Vitamin D3 Receptor (D2K6W) Rabbit mAb.免疫组织化学方法检测石蜡包埋的人乳腺癌组织,使用的抗体为Vitamin D3 Receptor (D2K6W) Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Vitamin D3 Receptor (D2K6W) Rabbit mAb.免疫组织化学方法检测石蜡包埋的人结肠癌组织,使用的抗体为Vitamin D3 Receptor (D2K6W) Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded mouse kidney, Vdr (+/+; left) or Vdr (-/-; right), using Vitamin D3 Receptor (D2K6W) Rabbit mAb (Tissues courtesy of Dr. Marie Demay, Massachusetts General Hospital).免疫组织化学方法检测石蜡包埋的小鼠肾脏组织,维生素D3受体(+/+; 左图)或维生素D3受体(-/-; 右图),使用的抗体为Vitamin D3 Receptor (D2K6W) Rabbit mAb(组织由麻省综合医院的Dr. Marie Demay友好提供)。

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Vitamin D3 Receptor (D2K6W) Rabbit mAb.Western blot方法检测多个细胞系提取物,使用的抗体为Vitamin D3 Receptor (D2K6W) Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of kidney tissue extracts from Vdr (+/+) and Vdr (-/-) mice using Vitamin D3 Receptor (D2K6W) Rabbit mAb (upper) and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). (Vdr (+/+) and Vdr (-/-) kidneys were kindly provided by Dr. Marie Demay, Massachusetts General Hospital).Western blot方法检测来自Vdr (+/+)和Vdr (-/-)小鼠肾脏组织提取物,使用的抗体为Vitamin D3 Receptor (D2K6W) Rabbit mAb (上图)和GAPDH (D16H11) XP® Rabbit mAb #5174 (下图).(Vdr (+/+) 和Vdr (-/-)肾脏由麻省综合医院的Dr. Marie Demay友好提供。)

Western Blotting

Western Blotting

Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with constructs expressing Myc/DDK-tagged full-length human vitamin D receptor isoform A (hVDRA-Myc/DDK; +), full-length human pregnane X receptor (hPXR-Myc/DDK; +), and full-length human constitutive androstane receptor (hCAR-Myc/DDK; +), using Vitamin D3 Receptor (D2K6W) Rabbit mAb (upper) or DYKDDDDK Tag Antibody #2368 (lower).Western blot方法检测293T细胞提取物,细胞不转染(-)或用构建表达Myc/DDK-标记的全长人维生素D3受体A亚型 (hVDRA-Myc/DDK; +),人全长孕烷受体(hPXR-Myc/DDK; +)和人全长组成型雄甾烷受体(hCAR-Myc/DDK; +),使用的抗体为Vitamin D3 Receptor (D2K6W) Rabbit mAb (上图) 或DYKDDDDK Tag Antibody #2368 (下图).

IP

IP

Immunoprecipitation of vitamin D3 receptor from T-47D cell extracts, using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or Vitamin D3 Receptor (D2K6W) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using Vitamin D3 Receptor (D2K6W) Rabbit mAb.免疫沉淀方法检测T-47D细胞提取物的维生素D3受体,使用的抗体为Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (泳道2) 或 Vitamin D3 Receptor (D2K6W) Rabbit mAb (泳道3). 泳道1是加入10%的蛋白。用 Vitamin D3 Receptor (D2K6W) Rabbit mAb进行Western blot分析。

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 LS180 cells treated with calcitriol (10nM, 3 hours) and either 10 μl of Vitamin D3 Receptor (D2K6W) 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 c-Fos Upstream Primers #25661, human UCA1 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

Although originally identified based on their roles in calcium and bone homeostasis, the vitamin D3 receptor (VDR/NR1I1) and its ligand 1-α, 25-dihydroxycholecalciferol [1α, 25(OH)2D3] are now recognized to exert biological effects in almost every tissue of the human body. Targets for vitamin D signaling include the central nervous system, skin, immune system, endocrine glands, kidney, and colon. At the cellular level, vitamin D signaling affects proliferation, differentiation, and apoptosis of both normal and transformed cells. Within the steroid receptor gene family, VDR belongs to the NR1I subfamily that also includes NR1I2/PXR and NR1I3/CAR. The human VDR gene is composed of 11 exons that encode six domains (A-F) of the full length VDR protein, which includes an N-terminal dual zinc finger DNA binding domain, a C-terminal ligand-binding activity domain, and an extensive unstructured region that links the two functional domains together (1). Upon 1α, 25(OH)2D3 binding to the hormone ligand-binding domain, VDR is stabilized by the phosphorylation of Ser51 in the DNA-binding domain by PKC (2), and Ser208 in the hinge region by casein kinase II (3). VDR associates with the retinoic acid receptor (RXR) through dimerization domains. The 1α, 25(OH)2D3-VDR-RXR complex binds to the vitamin D response elements (VDREs) in the promoters of target genes through the DNA-binding domain. Ligand-induced conformation changes in VDR results in the dissociation of the co-repressor, silencing-mediator for retinoid and thyroid hormone receptors (SMRT), and allows interaction of the VDR activation function (AF2) transactivation domain with transcriptional coactivators (1). 
 Studies have shown that variable VDR expression is associated with different forms or stages of cancer and likely results from tissue-type variation in 1α, 25(OH)2D3 signaling. In the case of colon cancer, research indicates that VDR expression is relatively higher in hyperplastic colon polyps and during early tumorigenesis but diminishes in later stage, poorly differentiated tumors. Multiple studies suggest that 1α, 25(OH)2D3 may be an attractive target for development as a therapeutic anticancer agent (4,5) .虽然最初根据维生素D3受体在钙和骨稳态中的作用对其进行认定,但现在在人体几乎每个组织中,维生素D3受体(VDR/NR1I1)及其配体1-α,25 - 二羟胆钙化醇[1α,25(OH)2D3]发挥的生物效应被识别。维生素D的信号靶标包括中枢神经系统、皮肤、免疫系统、内分泌腺体、肾和结肠。在细胞水平上,维生素D信号影响正常和转化细胞的增殖、分化和凋亡。类固醇受体基因家族中,VDR属于NR1I亚科,该亚科还包括NR1I2/PXR和NR1I3/CAR。人维生素D受体基因由编码全长VDR蛋白6个域(AF)的11个外显子组成,其中包括一个N-末端的双锌指结构DNA结合域,一个C-末端配体结合活性域和广泛链接这两个功能域的非结构化区域(1)。1α,25(OH)2D3结合激素配体结合域,由PKC磷酸化DNA结合结构域的丝氨酸(51位)(2)和酪蛋白激酶II磷酸化铰链区的丝氨酸(208位)(3),使VDR稳定。VDR与维甲酸受体(RXR)通过二聚化联合。 1α,25(OH)2D3 VDR-RXR复合物结合维生素D3响应元件(VDREs)通过DNA结合结构域启动靶基因。VDR中配体诱导的构象变化,导致辅阻遏物、维甲酸和甲状腺激素受体沉默介质(SMRT)的降解,并允许VDR活化功能转录激活域(AF2)与转录共激活因子的相互作用(1)。研究表明,可变VDR的表达与癌症不同类型或阶段有关,这可能是由1α, 25(OH)2D3信号中组织类型的变异造成的。研究表明,在结肠癌的情况下,VDR在增生性结肠息肉和早期肿瘤中表达相对较高,但在以后的阶段和低分化肿瘤中表达减少。多项研究表明,1α,25(OH)2D3可能是作为一种治疗型抗癌药物开发有吸引力的靶点(4,5)。

  1. Haussler, M.R. et al. (1998) J Bone Miner Res 13, 325-49.
  2. Hsieh, J.C. et al. (1991) Proc Natl Acad Sci U S A 88, 9315-9.
  3. Jurutka, P.W. et al. (1993) J Biol Chem 268, 6791-9.
  4. Matusiak, D. et al. (2005) Cancer Epidemiol Biomarkers Prev 14, 2370-6.
  5. Deeb, K.K. et al. (2007) Nat Rev Cancer 7, 684-700.

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