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#9955 4E-BP Antibody Sampler Kit

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2019年1月17日15時10分 現在
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4E-BP1 製品一覧

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キット内容 容量 用途 種交差性 検出分子量 アイソタイプ
Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb #2855 20 µl WB, IHC-P, IF-IC, F H, M, R, Mk, Dm 15 to 20 Rabbit IgG
Non-phospho-4E-BP1 (Thr46) (87D12) Rabbit mAb #4923 20 µl WB, IF-IC, F H, M, R, Mk 15-20 Rabbit IgG
Phospho-4E-BP1 (Ser65) Antibody #9451 20 µl WB, IP H, M, R, Mk 15 to 20 Rabbit
4E-BP1 (53H11) Rabbit mAb #9644 20 µl WB, IP, IHC-P, IF-IC, F H, M, R, Mk 15-20 Rabbit IgG
4E-BP2 Antibody #2845 20 µl WB, IP, IHC-P, F H, M, R, Mk, B 15 to 20 Rabbit
Phospho-4E-BP1 (Thr70) Antibody #9455 20 µl WB, IP H, M, R, Mk 15 to 20 Rabbit
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl WB Goat

Applications Key: W=Western Blotting, IHC-P=Immunohistochemistry (Paraffin), IF-IC=Immunofluorescence (Immunocytochemistry), F=Flow Cytometry, IP=Immunoprecipitation
Reactivity Key: H=Human, M=Mouse, R=Rat, Mk=Monkey, Dm=D. melanogaster, B=Bovine

貯法
-20℃
社内データ

Western Blotting

Western Blotting

Western blot analysis of extracts from 293 cells, untreated or treated with 20% FBS for the indicated number of minutes, using Phospho-4E-BP1 (Thr70) Antibody (upper) or 4E-BP1 Antibody #9452 (lower).

Western Blotting

Western Blotting

After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.

Western Blotting

Western Blotting

Western blot analysis of extracts from A673 cells, untreated or nocodazole-treated (100 ng/ml, 16 hrs), using 4E-BP2 Antibody (upper) or 4E-BP1 Antibody #9452 (lower). Extracts were treated with lambda phosphatase NEB#P0753 (10,000 U/ml for 1 hour) to dephosphorylate both proteins.


Western Blotting

Western Blotting

Western blot analysis of extracts from COS cells, treated with λ phosphatase or 20% FBS as indicated, using Non-phospho-4E-BP1 (Thr46) (87D12) Rabbit mAb (upper), Phospho-4E-BP1 (Thr37/46) Antibody #9459 (middle), and 4E-BP1 Antibody #9452 (lower).

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells using 4E-BP1 (53H11) Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using 4E-BP1 (53H11) Rabbit mAb.


Western Blotting

Western Blotting

Western blot analysis of extracts from 293T cells using 4E-BP1 Antibody #9452 (upper) and Phospho-4E-BP1 (Thr37/46) Antibody #2855 (lower). The cells were starved for 24 hours in serum-free medium and underwent a 1 hour amino acid deprivation. Amino acids were replenished for 1 hour. Cells were then either untreated (-) or treated with 100 nM insulin (+) for 30 minutes.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293 cells using 4E-BP1 Antibody #9644 (lower) and Phospho-4E-BP1 (Ser65) Antibody #9451 (upper). The cells were starved for 24 hours in serum-free medium and underwent a 1 hour amino acid deprivation. Amino acids were replenished for 1 hour. Cells were then either untreated (-) or treated with 100 nM insulin (+) for 30 minutes.

Western Blotting

Western Blotting

Western blot analysis of bacterially expressed GST-4E-BP1 and of extracts from NIH/3T3 cells, using 4E-BP2 Antibody and 4E-BP1 Antibody #9452.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of COS cells, untreated (blue) or λ phosphatase-treated (green), using Non-phospho-4E-BP1 (Thr46) (87D12) Rabbit mAb compared to a nonspecific negative control antibody (red).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using 4E-BP1 (53H11) Rabbit mAb in the presence of control peptide (left) or 4E-BP1 blocking peptide #1053 (right).


IF-IC

IF-IC

Confocal immunofluorescent analysis of 293 cells, expressing either nontargeting shRNA (left) or shRNA targeting 4E-BP1/2 (right), using 4E-BP1 (53H11) Rabbit mAb #9644 (green). Red = Propidium Iodide (PI)/RNase Staining Solution #4084.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma, showing cytoplasmic and nuclear localization, using 4E-BP2 Antibody.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human lymphoma using Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human hepatocellular carcinoma using 4E-BP1 (53H11) Rabbit mAb.

IF-IC

IF-IC

Confocal immunofluorescent analysis of 293 shRNA Scramble cells either serum starved (far left); serum starved and treated with U0126 #9903 (10 μM, 2 hr), LY294002 #9901 (50 μM, 2 hr), and Rapamycin #9904 (10 nM, 2 hr) (center, left); serum starved and treated with l phosphatase (10,000 U/mL, 2 hr) ) (center, right); and 293 shRNA 4E-BP1/2 KO treated with U0126 #9903 (10 μM, 2 hr), LY294002 #9901 (50 μM, 2 hr), and Rapamycin #9904 (10 nM, 2 hr) (far right), using Non-phospho-4E-BP1 (Thr46) (87D12) Rabbit mAb (green). Red = Propidium Iodide (PI)/RNase Staining Solution.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human lung carcinoma, using 4E-BP2 Antibody.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis using Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb on SignalSlide (TM) Phospho-Akt (Ser473) IHC Controls #8101 (paraffin-embedded LNCaP cells untreated (left) or LY294002-treated (right)).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human lung carcinoma using 4E-BP1 (53H11) Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human follicular carcinoma (thyroid), using 4E-BP2 Antibody.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb in the presence of control peptide (left) or Phospho-4E-BP1 (Thr37/46) Blocking Peptide #1052 (right).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded mouse spleen, 4E-BP1/2 wild type (left) or 4E-BP1 knockout (right), using 4E-BP1 (53H11) Rabbit mAb. 4E-BP wild type and knockout tissues kindly provided by Dr. Nahum Sonenberg, McGill University.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of HeLa cells, using 4E-BP2 Antibody (blue) compared to a nonspecifc negative control antibody (red).


Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Jurkat cells, untreated (green) or LY294002, Wortmannin and U0126-treated (blue), using Phospho-4E-BP1 (Thr36/46) (236B4) Rabbit mAb compared to a nonspecific negative control antibody (red).

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of COS cells using 4E-BP1 (53H11) Rabbit mAb (blue) compared to a nonspecific negative control antibody (red).

IF-IC

IF-IC

Confocal immunofluorescent analysis of 293 cells, expressing either non-targeting shRNA (top) or shRNA targeting 4E-BP1/2 (bottom), using Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb (green). To confirm phospho-specificity, cells were treated with an inhibitor cocktail consisting of LY294002 #9901, U0126 #9903, and Rapamycin #9904 (50 μM; 10 μm; 10 nM; 2 hr) (left), stimulated with insulin (100 nM, 30 min; middle), or processed with λ-phosphatase following insulin stimulation (right). Red = Propidium Iodide (PI)/RNase Staining Solution (#4087).


バックグラウンド

Translation repressor protein 4E-BP1 (also known as PHAS-1) inhibits cap-dependent translation by binding to the translation initiation factor eIF4E. Hyperphosphorylation of 4E-BP1 disrupts this interaction and results in activation of cap-dependent translation (1). Both the PI3 kinase/Akt pathway and FRAP/mTOR kinase regulate 4E-BP1 activity (2,3). Multiple 4E-BP1 residues are phosphorylated in vivo (4). While phosphorylation by FRAP/mTOR at Thr37 and Thr46 does not prevent the binding of 4E-BP1 to eIF4E, it is thought to prime 4E-BP1 for subsequent phosphorylation at Ser65 and Thr70 (5).

4E-BP2 and 4E-BP3 share high sequence homology with 4E-BP1, including conservation of the major FRAP/mTOR-dependent phosphorylation sites. Preliminary data suggests that phosphorylation of 4E-BP2 is regulated in a similar manner to that of 4E-BP1, although phosphorylation of this protein has not been as extensively studied (6).

使用例
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2498   eIF4G Antibody
6883   SignalFire™ ECL Reagent
7074   Anti-rabbit IgG, HRP-linked Antibody
7216   PathScan® Phospho-4E-BP1 (Thr37/Thr46) Sandwich ELISA Kit
7727   Biotinylated Protein Ladder Detection Pack
9452   4E-BP1 Antibody
9456   Phospho-4E-BP1 (Ser65) (174A9) Rabbit mAb
9459   Phospho-4E-BP1 (Thr37/46) Antibody
9741   Phospho-eIF4E (Ser209) Antibody
9742   eIF4E Antibody

DRAQ5 is a registered trademark of Biostatus Limited.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
LumiGLO is a registered trademark of Kirkegaard & Perry Laboratories.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.

本製品は試験研究用です。

4E-BP Antibody Sampler Kit

Immune Cell Signaling Pathways

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