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#13643 PKCθ (E1I7Y) Rabbit mAb

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希望納入価格 (円)
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2020年2月26日16時0分 現在
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#13643S100 μL60,000
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PKCTHETA 製品一覧

感度分子量 (kDa)抗体の由来貯法
内在性78Rabbit 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 ウシ
13643 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) (1:50)免疫蛍光細胞染色 (IF-IC) (1:400)フローサイトメトリー (1:400)

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PKCθタンパク質を検出します。
使用抗原
ヒトのPKCθタンパク質のPro632 周辺領域 (合成ペプチド)

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社内データ

※下記の社内データは、すべて13643 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 wild-type and PKCθ (-/-) mouse splenocytes using PKCθ (E1I7Y) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). Extracts from wild-type and PKCθ (-/-) mouse splenocytes were kindly provided by Dr. Morgan Huse (Memorial Sloan-Kettering Cancer Center).

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using PKCθ (E1I7Y) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).

IP

IP

Immunoprecipitation of PKCθ from Jurkat cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or PKCθ (E1I7Y) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using PKCθ (E1I7Y) Rabbit mAb.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon using PKCθ (E1I7Y) Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human gastrointestinal stromal tumor (GIST) using PKCθ (E1I7Y) Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human lymph node using PKCθ (E1I7Y) Rabbit mAb.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded Jurkat (positive, left) or RL7 (negative, right) cell pellets using PKCθ (E1I7Y) Rabbit mAb.

IF-IC

IF-IC

Confocal immunofluorescent analysis of Jurkat (positive, left) and Raji (negative, right) cells using PKCθ (E1I7Y) Rabbit mAb (green). Blue pseudocolor= DRAQ5® #4084 (fluorescent DNA dye).

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of mouse splenocytes using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (left) and PKCθ (E1I7Y) Rabbit mAb (right). Splenocytes were co-stained with anti-CD3 APC and the Anti-rabbit IgG (H+L), F(ab')2 Fragment (PE Conjugate) #8885 was used as a secondary antibody.


バックグラウンド

Activation of protein kinase C (PKC) is one of the earliest events in a cascade that controls a variety of cellular responses, including secretion, gene expression, proliferation, and muscle contraction (1,2). PKC isoforms belong to three groups based on calcium dependency and activators. Classical PKCs are calcium-dependent via their C2 domains and are activated by phosphatidylserine (PS), diacylglycerol (DAG), and phorbol esters (TPA, PMA) through their cysteine-rich C1 domains. Both novel and atypical PKCs are calcium-independent, but only novel PKCs are activated by PS, DAG, and phorbol esters (3-5). Members of these three PKC groups contain a pseudo-substrate or autoinhibitory domain that binds to substrate-binding sites in the catalytic domain to prevent activation in the absence of cofactors or activators. Control of PKC activity is regulated through three distinct phosphorylation events. Phosphorylation occurs in vivo at Thr500 in the activation loop, at Thr641 through autophosphorylation, and at the carboxy-terminal hydrophobic site Ser660 (2). Atypical PKC isoforms lack hydrophobic region phosphorylation, which correlates with the presence of glutamic acid rather than the serine or threonine residues found in more typical PKC isoforms. The enzyme PDK1 or a close relative is responsible for PKC activation. A recent addition to the PKC superfamily is PKCμ (PKD), which is regulated by DAG and TPA through its C1 domain. PKD is distinguished by the presence of a PH domain and by its unique substrate recognition and Golgi localization (6). PKC-related kinases (PRK) lack the C1 domain and do not respond to DAG or phorbol esters. Phosphatidylinositol lipids activate PRKs, and small Rho-family GTPases bind to the homology region 1 (HR1) to regulate PRK kinase activity (7).

PKCθ is a novel protein kinase C that is predominantly expressed in T cells (8). Recruitment of PKCθ to the immunological synapse following T cell receptor stimulation plays an important role in the activation and proliferation of conventional T cells (9). Conversely, PKCθ negatively regulates the suppressive function of regulatory T cells and is excluded from regulatory T cell immunological synapses (10).

使用例
 
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DRAQ5 is a registered trademark of Biostatus Limited.
Tween is a registered trademark of ICI Americas, Inc.
SignalStain is a trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

本製品は試験研究用です。

PKCθ (E1I7Y) Rabbit mAb

Metabolic Reprogramming in Disease

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