Cell Signaling Technology

Product Pathways - Nuclear Receptor Signaling

Progesterone Receptor A/B (D8Q2J) XP® Rabbit mAb #8757

No. Size Price
8757S 100 µl ( 10 western blots ) ¥3,580.00 现货查询 购买询价
8757T 20 µl ( 2 western blots ) ¥1,400.00 现货查询 购买询价
8757 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human, Endogenous 90 (PR-A), 118 (PR-B) Rabbit IgG
IP 1:50
IHC-P 1:1000
F 1:200
IF-IC 1:800
ChIP 1:100

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, IHC-P=Immunohistochemistry (Paraffin), F=Flow Cytometry, IF-IC=Immunofluorescence (Immunocytochemistry), ChIP=Chromatin IP,

Homology

Species predicted to react based on 100% sequence homology: Monkey,

Specificity / Sensitivity

Progesterone Receptor A/B (D8Q2J) XP® Rabbit mAb recognizes endogenous levels of total progesterone receptor A and B proteins. This antibody does not cross-react with either the glucocorticoid receptor or the mineralocorticoid receptor.Progesterone Receptor A/B (D8Q2J) XP® Rabbit mAb能够识别内源性水平的总孕酮受体A和B蛋白。该抗体不与糖皮质激素受体或盐皮质激素受体中的任何一个发生交叉反应。

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Tyr541 of human progesterone receptor protein.该单克隆抗体通过用合成肽免疫动物制备,该合成肽是人孕酮受体蛋白酪氨酸(541位)附近的残基。

IF-IC

IF-IC

Confocal immunofluorescent analysis of T-47D (PR positive, left) and MDA-MB-231 (PR negative, right) cells using Progesterone Receptor A/B (D8Q2J) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).激光共聚焦免疫荧光方法检测T-47D细胞 (孕酮受体阳性, 左图)和MDA-MB-231 细胞(孕酮受体阴性, 右图),使用的抗体为Progesterone Receptor A/B (D8Q2J) XP® Rabbit mAb (绿色).肌动蛋白丝用DY-554鬼笔环肽标记(红色)。

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded cell pellets, T-47D (high PR, left), MCF-7 (low PR, middle) and MDA-MB-231 (PR negative, right), using Progesterone Receptor A/B (D8Q2J) XP® Rabbit mAb.免疫组织化学方法检测石蜡包埋的细胞球,T-47D (高孕酮受体表达, 左图), MCF-7 (低孕酮受体表达, 中图)和MDA-MB-231 (孕酮受体阴性表达, 右图),使用的抗体为Progesterone Receptor A/B (D8Q2J) XP® Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human infiltrating ductal breast carcinoma using Progesterone Receptor A/B (D8Q2J) XP® Rabbit mAb.免疫组织化学方法检测石蜡包埋的人乳腺浸润性导管癌组织,使用的抗体为Progesterone Receptor A/B (D8Q2J) XP® Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from T-47D (PR positive) and MDA-MB-231 (PR negative) cells using Progesterone Receptor A/B (D8Q2J) XP® Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).Western blot方法检测T-47D细胞 (孕酮受体阳性)和MDA-MB-231 细胞(孕酮受体阴性)提取物,使用的抗体为Progesterone Receptor A/B (D8Q2J) XP® Rabbit mAb (上图)或GAPDH (D16H11) XP® Rabbit mAb #5174 (下图).

Western Blotting

Western Blotting

Western blot analysis of extracts from T-47D cells, grown for 48 hr in phenol red-free medium supplemented with 5% charcoal-stripped FBS and then treated with either a vehicle control (-) or promegestone (R5020, 100 nM, 16 hr; +), using Progesterone Receptor A/B (D8Q2J) XP® Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). Prolonged treatment of PR-expressing cells with R5020 is known to induce PR downregulation and hyperphosphorylation, which is reflected by slower migration on SDS-PAGE.Western blot方法检测T-47D细胞提取物,细胞在不含酚红,含5%活性碳/葡聚糖处理FBS中培养48小时,之后空白处理(-)或普美孕酮(R5020, 100 nM, 16 hr; +)处理,使用的抗体为Progesterone Receptor A/B (D8Q2J) XP® Rabbit mAb (上图) 或GAPDH (D16H11) XP® Rabbit mAb #5174 (下图).用R5020延长表达孕酮受体细胞的处理时间,已知R5020诱导孕酮受体下调和高度磷酸化,反应在SDS-PAGE上是迁移速率较慢。

Chromatin IP

Chromatin IP

T-47D cells were cultured in phenol red-free media supplemented with 5% charcoal-stripped FBS for 48 hr and then either untreated (left panel) or promegestone-treated (R5020, 10 nM, 1 hr; right panel). Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 cells and 5 µl of Progesterone Receptor A/B (D8Q2J) 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 FKBP51 Intron 5 Primers #7859, human E2F-1 proximal enhancer site #1 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.T-47D细胞培养在含5%活性炭/葡聚糖处理FBS的培养基中,培养48小时,之后细胞不处理(左面板)或用普美孕酮处理 (R5020, 10 nM, 1 hr;右面板)。通过交联染色质进行染色质免疫沉淀,交联染色质来自于4 x 106个细胞和5 µl Progesterone Receptor A/B (D8Q2J) XP® Rabbit mAb或2 µl Normal Rabbit IgG #2729,使用的试剂盒为SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003.通过real-time PCR 方法,对提取的DNA进行量化,使用的引物为SimpleChIP® Human FKBP51 Intron 5 Primers #7859, human E2F-1 proximal enhancer site #1 primers和SimpleChIP® Human α Satellite Repeat Primers #4486.在每个样品中的免疫沉淀的DNA量表示为相当于加入染色质总量,相当于一个。

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of MDA MB-231 cells (blue) and T47D cells using Progesterone REceptor A/B (D8Q2J) XP® Rabbit mAb. Anti-rabbit IgG (H+L), F(ab')2 Fragment (ALexa FLuor® 488 Conjugate) #4412 was used as a secondary antibody.

Background

Human progesterone receptor (PR) is expressed as two forms: the full length PR-B and the short form PR-A. PR-A lacks the first 164 amino acid residues of PR-B (1,2). Both PR-A and PR-B are ligand activated, but differ in their relative ability to activate target gene transcription (3,4). The activity of PR is regulated by phosphorylation; at least seven serine residues are phosphorylated in its amino-terminal domain. Three sites (Ser81, Ser102, and Ser162) are unique to full length PR-B, while other sites (Ser190, Ser294, Ser345, and Ser400) are shared by both isoforms (5). Phosphorylation of PR-B at Ser190 (equivalent to Ser26 of PR-A) is catalyzed by CDK2 (6). Mutation of Ser190 results in decreased activity of PR (7), suggesting that the phosphorylation at Ser190 may be critical to its biological function.人孕酮受体(PR)表现为两种形式:全长的孕酮受体B和短的孕酮受体A。PR A缺少PR B的第164个氨基酸残基(1,2)。PR A和PR B都是配体激活,但其激活靶基因转录的相对能力是不同的(3,4)。PR的活性受磷酸化调节。在其氨基末端区域,至少有七个丝氨酸残基被磷酸化。丝氨酸三个位点(81,102,162位)是全长PR特有的,而其他丝氨酸位点(190,294,345和400位)是两个亚型所共有的(5)。PR B丝氨酸(190位)(相当于PR A丝氨酸(26位))的磷酸化是由CDK2催化的(6)。丝氨酸(190位)的突变导致PR活性降低(7),这表明丝氨酸(190位)的磷酸化对其生物功能来说是至关重要的。

  1. Evans, R.M. (1988) Science 240, 889-895.
  2. Kastner, P. et al. (1990) EMBO J. 112, 1603-1614.
  3. Giangrande, P.H. et al. (2000) Mol. Cell. Biol. 20, 3102-3115.
  4. Wen, D.X. et al. (1994) Mol. Cell. Biol. 14, 8356-8364.
  5. Clemm, D.L. et al. (2000) Mol. Endocrinol. 14, 52-65.
  6. Zhang, Y. et al. (1997) Mol. Endocrinol. 11, 823-832.
  7. Takimoto, G.S. et al. (1996) J. Biol. Chem. 271, 13308-13316.

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