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#92570 Apoptosis/Necroptosis Antibody Sampler Kit

 
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希望納入価格 (円)
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2019年1月17日15時10分 現在
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Apoptosis 製品一覧

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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-RIP (Ser166) (D1L3S) Rabbit mAb #65746 20 µl WB H 78-82 Rabbit IgG
RIP (D94C12) XP® Rabbit mAb #3493 20 µl WB, IP, IF-IC, F H, M, R, Hm, Mk 78 Rabbit IgG
Phospho-MLKL (Ser358) (D6H3V) Rabbit mAb #91689 20 µl WB H 54 Rabbit IgG
MLKL (D2I6N) Rabbit mAb #14993 20 µl WB H 54 Rabbit IgG
Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb #9664 20 µl WB, IP, IHC-P, IHC-F, IF-IC, F H, M, R, Mk B, Dg, Pg 17, 19 Rabbit IgG
Caspase-3 (D3R6Y) Rabbit mAb #14220 20 µl WB H, M, R, Mk 35, 19, 17 Rabbit IgG
Cleaved Caspase-8 (Asp391) (18C8) Rabbit mAb #9496 20 µl WB, IHC-P, IF-IC, F H 18, 41, 43 Rabbit IgG
Caspase-8 (D35G2) Rabbit mAb #4790 20 µl WB H, M, R Pg 10, 57 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, IF-IC=Immunofluorescence (Immunocytochemistry), F=Flow Cytometry, IHC-P=Immunohistochemistry (Paraffin), IHC-F=Immunohistochemistry (Frozen)
Reactivity Key: H=Human, M=Mouse, R=Rat, Hm=Hamster, Mk=Monkey

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 C6 (rat), NIH/3T3 (mouse), and Jurkat (human) cells, untreated or treated with staurosporine #9953 (1uM, 3hrs) or etoposide #2200 (25uM, 5hrs) as indicated, using Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of HeLa cells transfected with the death receptors Fas and DR5, or treated with cycloheximide (CHX) and TNF-alpha using Cleaved Caspase-8 (Asp391) (18C8) Rabbit mAb (upper), or total Caspase-8 (1C12) Mouse mAb (lower).


Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using RIP (D94C12) XP® Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa, THP-1, and YB2/0 cell lines using Caspase-8 (D35G2) Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of various cell lines, untreated (-) or treated with Staurosporine #9953 (1 μM; 3 hr) or with Etoposide #2200 (25 μM, overnight), using Caspase-3 (D3R6Y) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). MCF7 cells are negative for caspase-3 expression.


Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using MLKL (D2I6N) Rabbit mAb. KARPAS cell Line source: Dr. Abraham Karpas at the University of Cambridge.

Western Blotting

Western Blotting

Western blot analysis of HT-29 cells, untreated (-) or treated with combinations of the following treatments as indicated: Z-VAD (20 μM, added 30 min prior to other compounds; +), human TNF-α (hTNF-α, 20 ng/ml, 7 hr; +), SM-164 (100 nM, 7 hr; +), and necrostatin-1 (Nec-1, 50 μM, 7 hr; +), using Phospho-RIP (Ser166) (D1L3S) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).

Western Blotting

Western Blotting

Western blot analysis of HT-29 cells, untreated (-), or treated with combinations of the following treatments as indicated: Z-VAD (20 μM, added 30 min prior to other compounds; +), human TNF-α (hTNF-α, 20 ng/ml, 7 hr; +), SM-164 (100 nM, 7 hr; +), and necrostatin-1 (Nec-1, 50 μM, 7 hr; +), using Phospho-MLKL (Ser358) (D6H3V) Rabbit mAb (upper), or MLKL (D2I6N) Rabbit mAb #14993 (lower).


IP

IP

Immunoprecipitation of extracts from Jurkat cells, untreated or etoposide-treated (25uM, 5hrs), using Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb. Western blot was performed using the same antibody.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon (chronic inflammation) using Cleaved Caspase-8 (Asp391) (18C8) Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, untransfected or transfected with human RIP construct, using RIP (D94C12) XP® Rabbit mAb.


Western Blotting

Western Blotting

Western blot analysis of extracts from CTLL-2 cells, untreated (-) or treated with Cycloheximide #2112 (CHX, 10 μg/ml, overnight) followed by Human Tumor Necrosis Factor-α (hTNF-α) #8902 (20 ng/ml, 4 hr; +), using Caspase-8 (D35G2) Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length human MLKL protein (hMLKL-Myc/DDK; +), using MLKL (D2I6N) Rabbit mAb (upper) and Myc-Tag (71D10) Rabbit mAb #2278 (lower).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded mouse embryo, using Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb in the presence of control peptide (left) or Cleaved Caspase-3 (Asp175) Blocking Peptide (#1050) (right).


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded HeLa cells, untreated (left) or Cycloheximide and TNFα-treated (right), using Cleaved Caspase-8 (Asp391) (18C8) Rabbit mAb.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of MCF7 cells using RIP (D94C12) XP® Rabbit mAb (blue) compared to Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (red).

Western Blotting

Western Blotting

Western blot analysis of 293T cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length human Caspase-8 protein (hCasp8-Myc/DDK; +), full-length mouse Caspase-8 protein (mCasp8; +), or full-length human Caspase-10 protein (hCasp10; +), using Caspase-8 (D35G2) Rabbit mAb.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis using Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb on SignalSlide® Cleaved Caspase-3 IHC Controls #8104 (paraffin-embedded Jurkat cells, untreated (left) or etoposide-treated (right)).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human Non-Hodgkin's lymphoma using Cleaved Caspase-8 (Asp391) (18C8) Rabbit mAb.

IF-IC

IF-IC

Confocal immunofluorescent analysis of OVCAR8 cells using RIP (D94C12) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical staining of paraffin-embedded mouse embryo, showing cytoplasmic localization in apoptotic cells, using Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Jurkat cells, untreated (blue) or etoposide-treated (green), using Cleaved Caspase-8 (Asp391) (18C8) Rabbit mAb compared to a nonspecific negative control antibody (red).

IHC-F (frozen)

IHC-F (frozen)

Immunohistochemical analysis of frozen H1650 xenograft, using Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb.


Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Jurkat cells, untreated (blue) or treated with etoposide #2200 (green), using Cleaved Caspase-3(Asp175) (5A1E) Rabbit mAb compared to a nonspecific negative control antibody (red).

IF-IC

IF-IC

Confocal immunofluorescent images of HT-29 cells, untreated (left) or Staurosporine #9953 treated (right) labeled with Cleaved Caspase-3 (Asp175) (5A1E) 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

Apoptosis is a regulated physiological process leading to cell death (1,2). Caspases, a family of cysteine acid proteases, are central regulators of apoptosis. Caspases are synthesized as inactive zymogens containing a pro-domain followed by large (p20) and small subunits (p10) that are proteolytically processed in a cascade of caspase activity. Initiator caspases (including 8, 9, 10, and 12) are closely coupled to proapoptotic signals. Once activated, these caspases cleave and activate downstream effector caspases (including 3, 6, and 7), which in turn cleave cytoskeletal and nuclear proteins like PARP, α-fodrin, DFF, and lamin A, and induce apoptosis. Cytochrome c released from mitochondria is coupled to the activation of caspase-9, a key initiator caspase. Apoptosis induced through the extrinsic mechanisms involving death receptors in the tumor necrosis factor receptor superfamily activates caspase-8. Activated caspase-8 cleaves and activates downstream effector caspases, such as caspase-1, -3, -6, and -7. Caspase-3 is a critical executioner of apoptosis, as it is either partially or totally responsible for the proteolytic cleavage of many key proteins, such as the nuclear enzyme poly (ADP-ribose) polymerase (PARP).

Necroptosis, a regulated pathway for necrotic cell death, is triggered by a number of inflammatory signals, including cytokines in the tumor necrosis factor (TNF) family, pathogen sensors such as toll-like receptors (TLRs), and ischemic injury (3,4). Necroptosis is negatively regulated by caspase-8 mediated apoptosis in which the kinase RIP/RIPK1 is cleaved (5). Furthermore, necroptosis is inhibited by a small molecule inhibitor of RIP, necrostatin-1 (Nec-1) (6). Research studies show that necroptosis contributes to a number of pathological conditions, and Nec-1 has been shown to provide neuroprotection in models such as ischemic brain injury (7). RIP is phosphorylated at several sites within the kinase domain that are sensitive to Nec-1, including Ser14, Ser15, Ser161, and Ser166 (8). Phosphorylation drives association with RIP3, which is required for necroptosis (9-11). Mixed lineage kinase domain-like protein (MLKL) is a pseudokinase that was identified as downstream target of RIP3 in the necroptosis pathway (12). During necroptosis RIP3 is phosphorylated at Ser227, which recruits MLKL and leads to its phosphorylation at Thr357 and Ser358 (12). Knockdown of MLKL through multiple mechanisms results in inhibition of necroptosis (13). While the precise mechanism for MLKL-induced necroptosis is unclear, some studies have shown that necroptosis leads to oligomerization of MLKL and translocation to the plasma membrane, where it effects membrane integrity (14-17).

使用例
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12630   SignalFire™ Plus ECL Reagent
12757   SignalFire™ Elite ECL Reagent
14220   Caspase-3 (D3R6Y) Rabbit mAb
14993   MLKL (D2I6N) Rabbit mAb
3493   RIP (D94C12) XP® Rabbit mAb
4790   Caspase-8 (D35G2) Rabbit mAb
65746   Phospho-RIP (Ser166) (D1L3S) Rabbit mAb
6883   SignalFire™ ECL Reagent
7074   Anti-rabbit IgG, HRP-linked Antibody
7727   Biotinylated Protein Ladder Detection Pack
91689   Phospho-MLKL (Ser358) (D6H3V) Rabbit mAb
9496   Cleaved Caspase-8 (Asp391) (18C8) Rabbit mAb
9664   Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb

Tween is a registered trademark of ICI Americas, Inc.
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。

Apoptosis/Necroptosis Antibody Sampler Kit

Immune Cell Signaling Pathways

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