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#10419 GITR (D5V7P) Rabbit mAb

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#10419S100 μL58,000
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TNFRSF18 製品一覧

感度分子量 (kDa)抗体の由来貯法
内在性25Rabbit 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 -
10419 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 (1:50)免疫組織染色 (パラフィン) (1:100)免疫蛍光細胞染色 (IF-IC) (1:400)フローサイトメトリー (1:200)Immunohistochemistry (Leica® Bond™) (1:100)

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
GITR (D5V7P) Rabbit mAb recognizes endogenous levels of total GITR protein.
使用抗原
Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the amino terminus of human GITR protein.

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社内データ

※下記の社内データは、すべて10419 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 HuT 102, human CD8+ T cells, and Jurkat cells using GITR (D5V7P) Rabbit mAb(upper), and β-Actin (D6A8) Rabbit mAb #8457 (lower). CD8+ T cells were purified from human blood and stimulated for 9 days using beads coated with CD3 and CD28 antibodies in the presence of human interleukin-2 (hIL-2) #8907 (6.7 ng/ml).

IP

IP

Immunoprecipitation of GITR protein from HuT 102 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is GITR (D5V7P) Rabbit mAb. Western blot analysis was performed using GITR (D5V7P) Rabbit mAb.

IHC-Leica® Bond™

IHC-Leica® Bond™

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using GITR (D5V7P) Rabbit mAb performed on the Leica® BOND™ Rx.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded HuT 102 cell pellet (left, positive) or Jurkat cell pellet (right, negative) using GITR (D5V7P) Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using GITR (D5V7P) Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human lung adenocarcinoma using GITR (D5V7P) Rabbit mAb.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human B-cell non-Hodgkin's lymphoma using GITR (D5V7P) Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human ovarian serous carcinoma using GITR (D5V7P) Rabbit mAb.

IF-IC

IF-IC

Confocal immunofluorescent analysis of HuT 102 (left, positive) and Jurkat (right, negative) cells using GITR (D5V7P) Rabbit mAb (green). Blue = Hoechst 33342 #4082 (fluorescent DNA dye).


Flow Cytometry

Flow Cytometry

Flow cytometric analysis of human peripheral blood mononuclear cells, untreated (left column) or treated with anti-CD3 (10 μg/ml, 72h) and anti-CD28 (5 μg/ml, 72h; right column), using GITR (D5V7P) Rabbit mAb (upper row) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype control #3900 (bottom row), and co-stained with a CD3 antibody. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody. Analysis was performed on CD3+ cells.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Jurkat cells (blue) and Hut102 cells (green), using GITR (D5V7P) Rabbit mAb (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

バックグラウンド

TNFRSF18, also known as glucocorticoid-induced tumor necrosis factor-receptor (TNFR)-related protein (GITR) and activation-inducible TNFR family receptor, encodes a type 1 membrane protein of the TNF-receptor superfamily (1). Three alternatively spliced transcript variants encoding distinct isoforms have been reported (2). GITR is an immune cell co-stimulatory receptor expressed constitutively at high levels on CD4+CD25+ T regulatory cells (Tregs), at low levels on naive and memory T cells, and is induced upon T cell activation (3-5). Studies show GITR can also be induced on NK cells, macrophages, and DCs (3, 4, 6). Although GITR does not have intrinsic enzymatic activity, TNFSF18 (also known as GITRL) expressed on antigen presenting cells binds to GITR resulting in recruitment of TNFR-associated factor family members and activation of the NF-kappa-B pathway in T cells (7). GITR ligation has been shown to play a role in CD8+ T cell activation, cytoxicity, and memory T cell survival (8-10). In the thymus, GITR is thought to play a key role in dominant immunological self-tolerance through thymic Treg differentiation and expansion (11). Of note, GITR ligation inhibits Treg suppressive function (12-13) and promotes effector T cell resistance to Treg suppression (14-15). Due to the combined effects on both Treg suppression and effector cell activation, GITR represents a unique opportunity for immunotherapeutic intervention in cancer (16).

使用例
 
関連製品
12630   SignalFire™ Plus ECL Reagent
12757   SignalFire™ Elite ECL Reagent
13079   Immunohistochemistry Application Solutions Kit (Rabbit)
13953   Prestained Protein Marker, Broad Range (11-190 kDa)
14166   Hematoxylin
14177   SignalStain® Mounting Medium
14746   SignalStain® Citrate Unmasking Solution (10X)
15019   Animal-Free Blocking Solution (5X)
6883   SignalFire™ ECL Reagent
7071   Phototope-HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
7074   Anti-rabbit IgG, HRP-linked Antibody
7727   Biotinylated Protein Ladder Detection Pack
8059   SignalStain® DAB Substrate Kit
8112   SignalStain® Antibody Diluent
8114   SignalStain® Boost IHC Detection Reagent (HRP, Rabbit)
9997   Tris Buffered Saline with Tween® 20 (TBST-10X)
9999   Nonfat Dry Milk

BOND is a trademark of Leica Biosystems Melbourne Pty. Ltd. No affiliation or sponsorship between CST and Leica Microsystems IR GmbH or Leica Biosystems Melbourne Pty. Ltd is implied.
Tween is a registered trademark of ICI Americas, Inc.
SignalStain is a trademark of Cell Signaling Technology, Inc.
LEICA is a registered trade​mark of Leica Microsystems IR GmbH.
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。

GITR (D5V7P) Rabbit mAb

Immune Cell Signaling Pathways

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