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#10396 Phospho-Akt Isoform Antibody Sampler Kit

 
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2019年1月17日15時10分 現在
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キット内容 容量 用途 種交差性 ホモロジー† 検出分子量 アイソタイプ
Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb #4060 20 µl WB, IP, IHC-P, IF-IC, F H, M, R, Hm, Mk, Dm, Z, B C, X, Dg, Pg 60 Rabbit IgG
Akt (pan) (C67E7) Rabbit mAb #4691 20 µl WB, IP, IHC-P, IF-IC, F H, M, R, Mk, Dm Pg 60 Rabbit IgG
Phospho-Akt1 (Ser473) (D7F10) XP® Rabbit mAb (Akt1 Specific) #9018 20 µl WB, IP, IF-IC, F H, M, R 60 Rabbit IgG
Akt1 (C73H10) Rabbit mAb #2938 20 µl WB, IP H, M, R, Mk 60 Rabbit IgG
Phospho-Akt2 (Ser474) (D3H2) Rabbit mAb (Akt2 Specific) #8599 20 µl WB, IP H, M, R 60 Rabbit IgG
Akt2 (D6G4) Rabbit mAb #3063 20 µl WB, IP H, M, R, Mk 60 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl WB Goat

†Species predicted to react based on 100% sequence homology.

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

貯法
-20℃
社内データ

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 PC-3 cells, untreated or LY294002/wortmannin-treated, and NIH/3T3 cells, serum-starved or PDGF-treated, using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb (upper) or Akt (pan) (C67E7) Rabbit mAb #4691 (lower).

Western Blotting

Western Blotting

Western blot analysis of recombinant Akt1, Akt2 and Akt3 proteins, and extracts from various cell lines, using Akt (pan) (C67E7) Rabbit mAb.


Western Blotting

Western Blotting

Western blot analysis of extracts from various cell types using Akt1 (C73H10) Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Akt2 (D6G4) Rabbit mAb (upper) and Akt1 (C73H10) Rabbit mAb #2938 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from Akt1 (-/-) mouse embryonic fibroblasts (MEF) or Akt2 (-/-) MEF, untreated (-) or treated with Human Platelet-Derived Growth Factor AA (hPDGF-AA) #8913 (100 ng/ml, 15 min; +), using Phospho-Akt2 (Ser474) (D3H2) Rabbit mAb (upper), Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb #4060 (middle), or Akt (pan) (C67E7) Rabbit mAb #4691 (lower).


Western Blotting

Western Blotting

Western blot analysis of extracts from LNCaP cells, transfected with a construct expressing Akt1 shRNA or Akt2 shRNA, uninduced (-) or doxycycline-induced for the indicated times, using Phospho-Akt1 (Ser473) (D7F10) XP® Rabbit mAb (Akt1 Specific) (top), Phospho-Akt2 (Ser474) (D3H2) Rabbit mAb (Akt2 Specific) #8599 (2nd from top), Akt1 (C73H10) Rabbit mAb #2938 (middle), Akt2 (D6G4) Rabbit mAb #3063 (2nd from bottom), or PI3 Kinase p85 (19H8) Rabbit mAb #4257 (bottom). (Data kindly provided by Drs. Rebecca Chin and Alex Toker, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, MA).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded MDA-MB-468 xenograft using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb (left) or PTEN (138G6) Rabbit mAb #9559 (right). Note the presence of P-Akt staining in the PTEN deficient MDA-MB-468 cells.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human melanoma using Akt (pan) (C67E7) Rabbit mAb.


Western Blotting

Western Blotting

Western blot analysis of extracts from Akt1 (lane 1), Akt2 (lane 2) and Akt3 (lane 3) knock-out mouse embryonic fibroblasts (MEFs) and matched wild-type MEFs (lane 4) using Akt1 (C73H10) Rabbit mAb (upper) and Akt2 Antibody #2962 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from LNCaP cells, transfected with a construct expressing Akt1 shRNA or Akt2 shRNA, uninduced (-) or doxycycline-induced for the indicated times, using Phospho-Akt1 (Ser473) (D7F10) XP® Rabbit mAb (Akt1 Specific) #9018 (top), Phospho-Akt2 (Ser474) (D3H2) Rabbit mAb (Akt2 Specific) (2nd from top), Akt1 (C73H10) Rabbit mAb #2938 (middle), Akt2 (D6G4) Rabbit mAb #3063 (2nd from bottom), or PI3 Kinase p85 (19H8) Rabbit mAb #4257 (bottom). (Data kindly provided by Drs. Rebecca Chin and Alex Toker, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, MA).

Western Blotting

Western Blotting

Western blot analysis of purified recombinant phospho-Akt1, phospho-Akt2 and phospho-Akt3 proteins using Phospho-Akt1 (Ser473) (D7F10) XP® Rabbit mAb (Akt1 Specific) (upper), Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb #4060 (middle) and Akt (pan) (C67E7) Rabbit mAb #4691 (lower).


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Akt (pan) (C67E7) Rabbit mAb in the presence of control peptide (left) or Akt (pan) Blocking Peptide #1085 (right).

Western Blotting

Western Blotting

Western blot analysis of recombinant Akt1, Akt2 and Akt3 proteins using Akt1 (C73H10) Rabbit mAb (upper) and Akt (pan) (11E7) Rabbit mAb #4685 (lower).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma comparing SignalStain® Antibody Diluent #8112 (left) to TBST/5% normal goat serum (right) using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb #4060.


Western Blotting

Western Blotting

Western blot analysis of extracts from Akt1 (-/-) mouse embryonic fibroblast (MEF) or Akt2 (-/-) MEF, untreated or stimulated with hPDGF #8913 (100 ng/ml, 15 min), using Phospho-Akt1 (Ser473) (D7F10) XP® Rabbit mAb (Akt1 Specific) (upper), Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb #4060 (middle), or Akt (pan) (C67E7) Rabbit mAb #4691 (lower).

Western Blotting

Western Blotting

Western blot analysis of purified recombinant phospho-Akt1, phospho-Akt2 and phospho-Akt3 proteins using Phospho-Akt2 (Ser474) (D3H2) Rabbit mAb (Akt2 Specific) (upper), Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb #4060 (middle) and Akt (pan) (C67E7) Rabbit mAb #4691 (lower).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis using Akt (pan) (C67E7) 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 breast carcinoma using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Akt1 (-/-) mouse embryonic fibroblast (MEF) cells (blue) or Akt2 (-/-) MEF cells (green), PDGF-treated (100 ng/mL, 15 min), using Phospho-Akt1 (Ser473) (D7F10) XP® Rabbit mAb (Akt1 Specific). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary Ab.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Jurkat cells using Akt (pan) (C67E7) Rabbit mAb (blue) compared to a nonspecific negative control antibody (red).


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb on SignalSlide® Phospho-Akt (Ser473) IHC Controls #8101 (paraffin-embedded LNCaP cells, untreated (left) or LY294002-treated (right)).

IF-IC

IF-IC

Confocal immunofluorescent analysis of Akt1 (-/-) mouse embryonic fibroblast (MEF) (left column), or Akt2 (-/-) MEF (right column) cells, stimulated with hPDGF #8913 (100 ng/ml, 15 min) (top row) or inhibited with LY294002 #9901 (bottom row), using Phospho-Akt1 (Ser473) (D7F10) XP® Rabbit mAb (Akt1 Specific) (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

IF-IC

IF-IC

Confocal immunofluorescent analysis of C2C12 cells, LY294002-treated (left) or insulin-treated (right), using Akt (pan) (C67E7) Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red). Blue pseudocolor = DRAQ5™ (fluorescent DNA dye).


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded PTEN heterozygous mutant mouse endometrium using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb. (Tissue section courtesy of Dr. Sabina Signoretti, Brigham and Women's Hospital, Harvard Medical School, Boston, MA.)

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded U-87MG xenograft, untreated (left) or lambda phosphatase-treated (right), using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb.


Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Jurkat cells, untreated (green) or treated with LY294002 #9901, wortmannin #9951 and U0126 #9903 (blue), using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb compared to a nonspecific negative control antibody (red).

IF-IC

IF-IC

Confocal immunofluorescent analysis of C2C12 cells, LY294002-treated (left) or insulin-treated (right), using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin #8953 (red). Blue pseudocolor = DRAQ5®#4084 (fluorescent DNA dye).

バックグラウンド

Akt, also referred to as PKB or Rac, plays a critical role in controlling survival and apoptosis (1-3). This protein kinase is activated by insulin and various growth and survival factors to function in a wortmannin-sensitive pathway involving PI3 kinase (2,3). Akt is activated by phospholipid binding and activation loop phosphorylation at Thr308 by PDK1 (4) and by phosphorylation within the carboxy terminus at Ser473. The previously elusive PDK2 responsible for phosphorylation of Akt at Ser473 has been identified as mammalian target of rapamycin (mTOR) in a rapamycin-insensitive complex with rictor and Sin1 (5,6). Akt promotes cell survival by inhibiting apoptosis through phosphorylation and inactivation of several targets, including Bad (7), forkhead transcription factors (8), c-Raf (9), and caspase-9. PTEN phosphatase is a major negative regulator of the PI3 kinase/Akt signaling pathway (10). LY294002 is a specific PI3 kinase inhibitor (11). Another essential Akt function is the regulation of glycogen synthesis through phosphorylation and inactivation of GSK-3α and β (12,13). Akt may also play a role in insulin stimulation of glucose transport (12). In addition to its role in survival and glycogen synthesis, Akt is involved in cell cycle regulation by preventing GSK-3β-mediated phosphorylation and degradation of cyclin D1 (14) and by negatively regulating the cyclin dependent kinase inhibitors p27 Kip1 (15) and p21 Waf1/Cip1 (16). Akt also plays a critical role in cell growth by directly phosphorylating mTOR in a rapamycin-sensitive complex containing raptor (17). More importantly, Akt phosphorylates and inactivates tuberin (TSC2), an inhibitor of mTOR within the mTOR-raptor complex (18,19).

There are three Akt isoforms (Akt1, Akt2 and Akt3) in mammals (20). Akt activation requires phosphorylation by mTORC2 at Ser473 of Akt1, Ser474 of Akt2, and Ser472 of Akt3 (20).

使用例
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関連製品
12630   SignalFire™ Plus ECL Reagent
12757   SignalFire™ Elite ECL Reagent
2938   Akt1 (C73H10) Rabbit mAb
3063   Akt2 (D6G4) Rabbit mAb
4060   Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb
4691   Akt (pan) (C67E7) Rabbit mAb
6883   SignalFire™ ECL Reagent
7074   Anti-rabbit IgG, HRP-linked Antibody
7727   Biotinylated Protein Ladder Detection Pack
8599   Phospho-Akt2 (Ser474) (D3H2) Rabbit mAb (Akt2 Specific)
9018   Phospho-Akt1 (Ser473) (D7F10) XP® Rabbit mAb (Akt1 Specific)
9997   Tris Buffered Saline with Tween® 20 (TBST-10X)
9999   Nonfat Dry Milk

DRAQ5 is a registered trademark of Biostatus Limited.
SignalStain is a trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
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。

Phospho-Akt Isoform Antibody Sampler Kit

Immune Cell Signaling Pathways

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