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#9911 Phospho-Erk1/2 Pathway Sampler Kit

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2019年1月18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-c-Raf (Ser338) (56A6) Rabbit mAb #9427 20 µl WB H, M, R, Mk 74 Rabbit IgG
Phospho-MEK1/2 (Ser217/221) (41G9) Rabbit mAb #9154 20 µl WB, IP H, M, R, Mk C 45 Rabbit
Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb #4370 20 µl WB, IP, IHC-P, IF-IC, F H, M, R, Hm, Mk, Mi, Dm, Z, B, Dg, Pg, Sc C, Ce 44, 42 Rabbit IgG
Phospho-p90RSK (Ser380) (D3H11) Rabbit mAb #11989 20 µl WB, IHC-P, IF-IC H, M, R, Mk, Mi C, X, Z, B, Dg, Pg, Hr 90 Rabbit IgG
Phospho-MSK1 (Thr581) Antibody #9595 20 µl WB, IP, IHC-P H, M 90 Rabbit
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, Mk=Monkey, Hm=Hamster, Mi=Mink, Dm=D. melanogaster, Z=Zebrafish, B=Bovine, Dg=Dog, Pg=Pig, Sc=S. cerevisiae

貯法
-20℃
社内データ

Western Blotting

Western Blotting

Western blot analysis of extracts from 293 cells, transfected with wt MSK1 or mutant MSK1, untreated or TPA-treated (200 nM), using Phospho-MSK1 (Thr581) Antibody.

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 NIH3T3, HeLa and COS cells, untreated or treated with TPA, using Phospho-c-Raf (Ser338) (56A6) Rabbit mAb.


Western Blotting

Western Blotting

Western blot analysis of extracts from untreated or TPA-treated HeLa and NIH/3T3 cells, using Phospho-MEK1/2 (Ser217/221) (41G9) Rabbit mAb (upper), or MEK1/2 Antibody #9122 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from COS cells, untreated or treated with either U0126 #9903 (10 µM for 1h) or TPA #4174 (200 nM for 10 m), using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb #4370 and p44/42 MAPK (Erk1/2) (3A7) Mouse mAb #9107.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines, starved overnight and either untreated (-) or treated (+) with TPA #4174 (200 nM, 15 min), Human Epidermal Growth Factor (hEGF) #8916 (100 ng/mL, 15 min), or Human Platelet-Derived Growth Factor BB (hPDGF-BB) #8912 (100 ng/mL, 15 min) as indicated, using Phospho-p90RSK (Ser380) (D3H11) Rabbit mAb.


Western Blotting

Western Blotting

Western blot analysis of extracts from 293 cells, untreated, UV-treated or calyculin A-treated, using Phospho-MSK1 (Thr581) Antibody.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293, NIH/3T3 and C6 cells, treated with λ phosphatase or TPA #4174 as indicated, using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb (upper), or p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb #4695 (lower).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma, control (left) or λ phosphatase-treated (right), using Phospho-p90RSK (Ser380) (D3H11) Rabbit mAb.


Western Blotting

Western Blotting

Western blot analysis of extracts from 293 cells untreated or treated with UV and HeLa cells untreated or treated with calyculin A, using Phospho-MSK1 (Thr581) Antibody.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human ovarian carcinoma using Phospho-p90RSK (Ser380) (D3H11) Rabbit mAb.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, showing nuclear localization, using Phospho-MSK1 (Thr581) Antibody.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human lung carcinoma, untreated (left) or λ phosphatase-treated (right), using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded HeLa cell pellets, untreated (left) or treated with TPA #4174 (right), using Phospho-p90RSK (Ser380) (D3H11) Rabbit mAb.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb on SignalSlide™ Phospho-p44/42 MAPK (Thr202/Tyr204) IHC Controls #8103 (paraffin-embedded NIH/3T3 cells, treated with U0126 #9903 (left) or TPA #4174 (right).

IF-IC

IF-IC

Confocal immunofluorescent analysis of NIH/3T3 cells, serum-starved (left), treated with TPA #4174 (200 nM, 15 min; center), or treated with TPA followed by λ phosphatase (right), using Phospho-p90RSK (Ser380) (D3H11) Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Jurkat cells, treated with U0126 (10 µM, 2 hrs; green) or treated with TPA #4174 (200 nM, 30 min; blue) using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® 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.


IF-IC

IF-IC

Confocal immunofluorescent analysis of Drosophila egg chambers, untreated (top) or λ phosphatase-treated (bottom), using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb #4370 (green) and S6 Ribosomal Protein (54D2) Mouse mAb #2317 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

IF-IC

IF-IC

Confocal immunofluorescent analysis of HT1080 cells, starved overnight then treated with U0126 #9903 (10 uM, 2 h; left) or PDBu (Phorbol 12,13-Dibutyrate) #12808 (100 nM, 15 m; right) using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb #4370 (green) and β-Actin (8H10D10) Mouse mAb #3700 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

バックグラウンド

Mitogen-activated protein kinases (MAPKs) are a widely conserved family of serine/threonine protein kinases involved in many cellular programs, such as cell proliferation, differentiation, motility, and death. The p44/42 MAPK (Erk1/2) signaling pathway can be activated in response to a diverse range of extracellular stimuli including mitogens, growth factors, and cytokines (1-3), and research investigators consider it an important target in the diagnosis and treatment of cancer (4). Upon stimulation, a sequential three-part protein kinase cascade is initiated, consisting of a MAP kinase kinase kinase (MAPKKK or MAP3K), a MAP kinase kinase (MAPKK or MAP2K), and a MAP kinase (MAPK). Multiple p44/42 MAP3Ks have been identified, including members of the Raf family, as well as Mos and Tpl2/COT. MEK1 and MEK2 are the primary MAPKKs in this pathway (5,6). MEK1 and MEK2 activate p44 and p42 through phosphorylation of activation loop residues Thr202/Tyr204 and Thr185/Tyr187, respectively. Several downstream targets of p44/42 have been identified, including p90RSK (7) and the transcription factor Elk-1 (8,9). p44/42 are negatively regulated by a family of dual-specificity (Thr/Tyr) MAPK phosphatases, known as DUSPs or MKPs (10), along with MEK inhibitors, such as U0126 and PD98059.

使用文献
 
関連製品
12630   SignalFire™ Plus ECL Reagent
12757   SignalFire™ Elite ECL Reagent
4695   p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb
6883   SignalFire™ ECL Reagent
7074   Anti-rabbit IgG, HRP-linked Antibody
7727   Biotinylated Protein Ladder Detection Pack
9126   MEK1/2 (47E6) Rabbit mAb
9160   MEK1/2 Control Cell Extracts
9355   RSK1/RSK2/RSK3 (32D7) Rabbit mAb
9422   c-Raf Antibody

DRAQ5 is a registered trademark of Biostatus Limited.
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-Erk1/2 Pathway Sampler Kit

Immune Cell Signaling Pathways

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