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#48799 Mitochondrial Dynamics Antibody Sampler Kit

 
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2019年1月17日11時55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
Tom20 (D8T4N) Rabbit mAb #42406 20 µl WB, IP, IHC-P, IF-IC H, M, R, Mk 16 Rabbit IgG
OPA1 (D6U6N) Rabbit mAb #80471 20 µl WB, IP H, M, R 80-100 Rabbit IgG
Phospho-DRP1 (Ser616) (D9A1) Rabbit mAb #4494 20 µl WB, IF-IC, F H M, R 78-82 Rabbit IgG
DRP1 (D6C7) Rabbit mAb #8570 20 µl WB, IP, IF-IC H, M, R, Mk Pg 78-82 Rabbit IgG
Phospho-MFF (Ser146) Antibody #49281 20 µl WB H, M, R 25, 27 Rabbit
MFF (E5W4M) XP® Rabbit mAb #84580 20 µl WB, IP, IF-IC H, M, R B, Dg 25, 27, 30, 35 Rabbit IgG
Mitofusin-1 (D6E2S) Rabbit mAb #14739 20 µl WB, IP, IF-IC H 82 Rabbit IgG
Mitofusin-2 (D1E9) Rabbit mAb #11925 20 µl WB, IP, IF-IC H, Hm, Mk 80 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, Mk=Monkey, Hm=Hamster

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 various cell lines using DRP1 (D6C7) Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, untreated (-) or treated (+) with nocodazole (100 ng/ml, 17 hr), using Phospho-DRP1 (Ser616) (D9A1) Rabbit mAb (upper) and DRP1 (D6C7) Rabbit mAb #8570 (lower).


IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells using Mitofusin-2 (D1E9) Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Mitofusin-2 (D1E9) Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Mitofusin-1 (D6E2S) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).


Western Blotting

Western Blotting

Western blot analysis of extracts of MEFs from wild-type (-) and OPA knockout (OPA-/-; +) mice using OPA1 (D6U6N) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from MCF7 and HeLa cells, untreated (-) or treated with carbonyl cyanide 3-chlorophenylhydrazone (CCCP; +) using OPA1 (D6U6N) Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa and Saos-2 cells using Tom20 (D8T4N) Rabbit mAb (upper) and β-Tubulin (D2N5G) Rabbit mAb #15115 (lower).


Western Blotting

Western Blotting

Western blot analysis of extracts from MCF7 cells, untreated (-) or treated with CCCP (100μM, 2hr; +), using phospho-MFF (Ser146) Antibody (upper) or total MFF antibody (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with siRNA targeting human MFF (+), using MFF (E5W4M) XP® Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).

IP

IP

Immunoprecipitation of extracts from HCT 116 cells using DRP1 (D6C7) Rabbit mAb. The western blot was probed using the same antibody. Lane 1 is 10% input.


Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Jurkat cells, untreated (left) or λ phosphatase-treated (right), using Phospho-DRP1 (Ser616) (D9A1) Rabbit mAb and propidium iodide (DNA content).

IP

IP

Immunoprecipitation of Mitofusin-2 from PANC-1 cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or Mitofusin-2 (D1E9) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using Mitofusin-2 (D2D10) Rabbit mAb #9482.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, transfected with SignalSilence® Control siRNA (unconjugated) #6568 (-), or SignalSilence® Mitofusin-1 siRNA I #13274 (+), using Mitofusin-1 (D6E2S) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). The Mitofusin-1 (D6E2S) Rabbit mAb confirms silencing of mitofusin-1 expression, while the β-Actin (D6A8) Rabbit mAb is used as a loading control.


Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® OPA1 siRNA I (+), or SignalSilence® OPA1 siRNA II using OPA1 (D6U6N) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). The OPA1 (D6U6N) Rabbit mAb confirms silencing of OPA1 expression, while the β-Actin (D6A8) Rabbit mAb is used as a loading control.

Western Blotting

Western Blotting

Western blot analysis of extracts from MCF7 cells, untreated (-) or treated with CCCP (100uM, 2hr; +), using Phospho-MFF (Ser146) Antibody. The phospho-specificity of the antibody was verified by treating the membrane with (+) or without (-) calf intestinal phosphatase (CIP) after protein transfer.

Western Blotting

Western Blotting

Western blot analysis of extracts from wild-type mouse embryonic fibroblasts (MEFs) (WT), Mff (-/-) MEFs (Mff KO), or Mff (-/-) MEFs reconstituted with a construct expressing human MFF protein, isoform 5 (Mff KO+hMFF) using MFF (E5W4M) XP® Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). Lysates courtesy of Sebastien Herzig and Portia Lombardo, Reuben Shaw lab, Salk Institute.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded HeLa cell pellet (left, high-expressing) or Saos-2 cell pellet (right, low-expressing) using Tom20 (D8T4N) Rabbit mAb.

IF-IC

IF-IC

Confocal immunofluorescent analysis of ACHN cells using DRP1 (D6C7) Rabbit mAb (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 HeLa cells, untreated (left) or λ phosphatase-treated (right), using Phospho-DRP1 (Ser616) (D9A1) Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).


IP

IP

Immunoprecipitation of mitofusin-1 protein from HeLa cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or Mitofusin-1 (D6E2S) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using Mitofusin-1 (D6E2S) Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using MFF (E5W4M) XP® Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded mouse kidney using Tom20 (D8T4N) Rabbit mAb.


IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells using Mitofusin-1 (D6E2S) Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using OPA1 (D6U6N) Rabbit mAb.

IP

IP

Immunoprecipitation of MFF from HeLa cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is MFF (E5W4M) XP® Rabbit mAb. Western blot analysis was performed using MFF (E5W4M) XP® Rabbit mAb. Mouse Anti-Rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 was used for detection to avoid cross-reactivity with IgG.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human lung adenocarcinoma using Tom20 (D8T4N) Rabbit mAb.

IP

IP

Immunoprecipitation of OPA1 from MCF7 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is OPA1 (D6U6N) Rabbit mAb. Western blot analysis was performed using OPA1 (D6U6N) Rabbit mAb. A conformation specific secondary antibody was used to avoid reactivity with IgG.

IF-IC

IF-IC

Confocal immunofluorescent analysis of wild-type mouse embryonic fibroblasts (MEFs) (WT, left), Mff (-/-) MEFs (Mff KO, center), or Mff (-/-) MEFs reconstituted with a construct expressing human MFF protein, isoform 5 (Mff KO+hMFF, right) using MFF (E5W4M) XP® Rabbit mAb (green). Nuclei labeled with DAPI (blue). Image courtesy of Sebastien Herzig and Portia Lombardo, Reuben Shaw lab, Salk Institute.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human prostate carcinoma using Tom20 (D8T4N) Rabbit mAb.

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells using MFF (E5W4M) XP® Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded mouse small intestine using Tom20 (D8T4N) Rabbit mAb.


IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa (high expression, left) and Saos-2 (low expression, right) cells, using Tom20 (D8T4N) Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

バックグラウンド

Import of proteins into the mitochondria is regulated by the translocase of the outer mitochondrial membrane (TOM) complex, which facilitates transport through the outer mitochondrial membrane, and a complementary translocase of the inner membrane (TIM) complex, responsible for protein transport to the mitochondrial matrix. The TOM complex consists of the receptors Tom20, Tom22, and Tom70, and the channel-forming protein Tom40 (1). Tom20 is localized in the outer mitochondrial membrane and initially recognizes precursors with a presequence to facilitate protein import across the outer mitochondrial membrane (2).

Changes in mitochondrial dynamics regulated by environmental cues affect mitochondrial size and shape and have been shown to dramatically impact mitochondrial metabolism, apoptosis, and autophagy (3). These processes are largely controlled by mitochondrial dynamin-related GTPases, including mitofusin-1, mitofusin-2, OPA1, and DRP1. DRP1 regulates mitochondrial fission, while the mitofusins and OPA1 control fusion at the outer and inner mitochondrial membrane, respectively. These proteins are tightly regulated. OPA1 activity is regulated through alternative splicing and post-translational modifications, including complex proteolytic processing by multiple proteases (4-9). In addition, OPA1 expression can be induced under conditions of metabolic demand through a pathway involving Parkin induced NF-κB activation (10). DRP1 is regulated in part through multiple phosphorylation sites (11). Phosphorylation of DRP1 at Ser616 by MAPK or during mitosis by CDKs stimulates mitochondrial fission (12-14). Mitochondrial fission factor (MFF) is a tail-anchored protein that resides within the outer mitochondrial membrane and is part of the mitochondrial fission complex. MFF participates in mitochondrial fission by serving as one of multiple receptors for the GTPase dynamin-related protein 1 (Drp1) (15-18). AMPK directly phosphorylates MFF at two sites to allow for enhanced recruitment of Drp1 to the mitochondria (19). 

使用例
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関連製品
11925   Mitofusin-2 (D1E9) Rabbit mAb
14739   Mitofusin-1 (D6E2S) Rabbit mAb
42406   Tom20 (D8T4N) Rabbit mAb
4494   Phospho-DRP1 (Ser616) (D9A1) Rabbit mAb
49281   Phospho-MFF (Ser146) Antibody
7003   20X LumiGLO® Reagent and 20X Peroxide
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
80471   OPA1 (D6U6N) Rabbit mAb
84580   MFF (E5W4M) XP® Rabbit mAb
8570   DRP1 (D6C7) Rabbit mAb

DRAQ5 is a registered trademark of Biostatus Limited.
Tween is a registered trademark of ICI Americas, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
LumiGLO is a registered trademark of Kirkegaard & Perry Laboratories.
SignalSilence is a registered trademark of Cell Signaling Technology, Inc.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
DyLight is a trademark of Thermo Fisher Scientific, Inc. and its subsidiaries.

本製品は試験研究用です。

Mitochondrial Dynamics Antibody Sampler Kit

Metabolic Reprogramming in Disease

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