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#12041 Glucocorticoid Receptor (D6H2L) XP® Rabbit mAb

 
CSTコード 包装
希望納入価格 (円)
国内在庫 i
2019年11月15日15時30分 現在
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#12041S100 μL62,000
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#12041T20 μL39,000
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GLUCOCORTICOID RECEPTOR 製品一覧

感度分子量 (kDa)抗体の由来貯法
内在性94, 91Rabbit IgG-20℃
種交差性 (社内試験済)
交差する可能性がある種 i

社内試験はしていませんが、配列が100%相同であるため反応すると推定される種

ヒト、マウス、ラット、サル -
12041 の推奨プロトコール i

最適な結果を得るために:Cell Signaling Technology (CST) 社は、各製品の推奨プロトコールを使用することを強くお薦めいたします。
推奨プロトコールはCST社内試験の徹底的なバリデーションに基づいて作成されておりますので、正確かつ再現性の高い結果が得られます。

注:各製品に最適化されたプロトコールをリンクしています。

 

ウェスタンブロッティング (1:1000)免疫沈降 (1:100)免疫組織染色 (パラフィン) (1:400)免疫蛍光細胞染色 (IF-IC) (1:50)フローサイトメトリー (1:200)クロマチン免疫沈降 (1:50)クロマチン免疫沈降シークエンス (1:50)

下記ステップについては、データシートの右側もあわせてご参照ください。

IHC-P: 抗体希釈液 / 抗原賦活化

CST推奨プロトコールに欠かせない関連製品

特異性・感度
内在性レベルのGlucocorticoid Receptor (GR) タンパク質を検出します。GR-αやGR-βタンパク質とは交差しますが、Mineralocorticoid Receptor タンパク質とは交差しません。
使用抗原
ヒトのGR タンパク質のN末端領域 (組換えタンパク質)

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社内データ

※下記の社内データは、すべて12041 の推奨プロトコールで実験した結果です。

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Glucocorticoid Receptor (D6H2L) XP® Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293T cells, either mock transfected (-) or transfected with constructs expressing Myc/DDK-tagged full-length human glucocorticoid receptor-α (hGRα-Myc/DDK; +) or Myc/DDK-tagged full-length human mineralocorticoid receptor (hMR-Myc/DDK; +), using Glucocorticoid Receptor (D6H2L) XP® Rabbit mAb (upper) or DYKDDDDK Tag Antibody #2368 (lower).

IP

IP

Immunoprecipitation of glucocorticoid receptor from HeLa cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or Glucocorticoid Receptor (D6H2L) XP® Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using Glucocorticoid Receptor (D6H2L) XP® Rabbit mAb.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Glucocorticoid Receptor (D6H2L) XP® Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Glucocorticoid Receptor (D6H2L) XP® Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded mouse stomach using Glucocorticoid Receptor (D6H2L) XP® Rabbit mAb.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded HeLa cell pellets, untreated (left) or dexamethasone-treated (right), using Glucocorticoid Receptor (D6H2L) XP® Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human prostate carcinoma using Glucocorticoid Receptor (D6H2L) XP® Rabbit mAb.

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells, grown in phenol red-free media containing 5% charcoal-stripped FBS for 2 d and either untreated (left) or treated with dexamethasone (100 nM, 2 hr; right), using Glucocorticoid Receptor (D6H2L) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).


Flow Cytometry

Flow Cytometry

Human whole blood was fixed, lysed, and permeabilized as per the Cell Signaling Technology Flow Cytometry (Alternate) Protocol, and stained with CD3-PE, CD19-APC, and Glucocorticoid Receptor (D6H2L) XP® Rabbit mAb. B cell (green) and T cell (blue) population gates (left) were applied to a histogram depicting the mean fluorescence intensity of glucocorticoid receptor, compared to a nonspecific negative control antibody (red; right). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

Chromatin IP-seq

Chromatin IP-seq

Chromatin immunoprecipitations were performed with cross-linked chromatin from A549 cells cultured in media with 5% charcoal-stripped FBS for 3 d and then treated with 100 nM dexamethasone for 1 hr and Glucocorticoid Receptor (D6H2L) XP® Rabbit mAb, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figure shows binding across SLC19A2, a known target gene of GR (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product data sheet.

Chromatin IP

Chromatin IP

A549 cells were cultured in media with 5% charcoal-stripped FBS for 3 d and then either untreated (left panel) or dexamethasone-treated (100 nM, 1 hr; right panel). Chromatin immunoprecipitations were performed with cross-linked chromatin from A549 cells and Glucocorticoid Receptor (D6H2L) 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 SLC19A2 Promoter Primers #7681, human MT2A 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.


バックグラウンド

Glucocorticoid hormones control cellular proliferation, inflammation, and metabolism through their association with the glucocorticoid receptor (GR)/NR3C1, a member of the nuclear hormone receptor superfamily of transcription factors (1). GR is composed of several conserved structural elements, including a carboxy-terminal ligand-binding domain (which also contains residues critical for receptor dimerization and hormone-dependent gene transactivation), a neighboring hinge region containing nuclear localization signals, a central zinc-finger-containing DNA-binding domain, and an amino-terminal variable region that participates in ligand-independent gene transcription. In the absence of hormone, a significant population of GR is localized to the cytoplasm in an inactive form via its association with regulatory chaperone proteins, such as HSP90, HSP70, and FKBP52. On hormone binding, GR is released from the chaperone complex and translocates to the nucleus as a dimer to associate with specific DNA sequences termed glucocorticoid response elements (GREs), thereby enhancing or repressing transcription of specific target genes (2). It was demonstrated that GR-mediated transcriptional activation is modulated by phosphorylation (3-5). Although GR can be basally phosphorylated in the absence of hormone, it becomes hyperphosphorylated upon binding receptor agonists. It has been suggested that hormone-dependent phosphorylation of GR may determine target promoter specificity, cofactor interaction, strength and duration of receptor signaling, receptor stability, and receptor subcellular localization (3).

使用例
 
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Illumina is a registered trademark of Illumina, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

本製品は試験研究用です。

Glucocorticoid Receptor (D6H2L) XP® Rabbit mAb

Immune Cell Signaling Pathways

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