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#12675 Initiator Caspases 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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キット内容 容量 用途 種交差性 検出分子量 アイソタイプ
Cleaved Caspase-9 (Asp330) (E5Z7N) Rabbit mAb #52873 20 µl WB, IF-IC H 37 Rabbit IgG
Caspase-3 (8G10) Rabbit mAb #9665 20 µl WB, IP H, M, R, Mk 17, 19, 35 Rabbit IgG
Caspase-2 (C2) Mouse mAb #2224 20 µl WB H 12, 14, 48 Mouse IgG1
Caspase-9 (C9) Mouse mAb #9508 20 µl WB H, M, R, Hm, Mk 47/37/35 (H). 49/39/37 (M). 51/40/38 (R). Mouse IgG1
Caspase-8 (1C12) Mouse mAb #9746 20 µl WB, IP H 18, 43, 57 Mouse IgG1
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl WB Goat
Anti-mouse IgG, HRP-linked Antibody #7076 100 µl WB Horse

Applications Key: W=Western Blotting, IF-IC=Immunofluorescence (Immunocytochemistry), IP=Immunoprecipitation
Reactivity Key: H=Human, M=Mouse, R=Rat, Mk=Monkey, Hm=Hamster

貯法
-20℃
社内データ

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells, untreated (left) or treated with Staurosporine #9953 (1 μM, 3 hr; right), using Cleaved Caspase-9 (Asp330) (E5Z7N) 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 SKW6.4 cells, untreated or anti-Fas-treated (1 µg/ml), and Jurkat cells, untreated or etoposide-treated (25 µM), using Caspase-8 (1C12) Mouse mAb.

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 Jurkat cells, untreated or etoposide-treated (5 hr, 25μM), using Caspase-2 (C2) Mouse mAb.


Western Blotting

Western Blotting

Western blot analysis of extracts from Jurkat cells (human), L929 cells (mouse), and C6 cells (rat), untreated or treated with staurosporine or cytochrome c as indicated, using Caspase 9 (C9) Mouse mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 (-) or SignalSilence® Caspase-3 siRNA II (+), using Caspase-3 (8G10) Rabbit mAb and α-Tubulin (11H10) Rabbit mAb #2125. Caspase-3 (8G10) Rabbit mAb confirms silencing of caspase-3 expression, while the α-Tubulin (11H10) Rabbit mAb is used to control for loading and specificity of caspase-3 siRNA.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa and Jurkat cells, untreated (-) or treated with Staurosporine #9953 (1 μM, 3 hr; +) or Etoposide #2200 (25 μM, overnight; +), using Cleaved Caspase-9 (Asp330) (E5Z7N) Rabbit mAb (upper), Caspase-9 (C9) Mouse mAb #9508 (middle), or β-Actin (D6A8) Rabbit mAb #8457 (lower).


Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 (-) or SignalSilence® Caspase-3 siRNA I (+), using Caspase-3 (8G10) Rabbit mAb and p42 MAPK Antibody #9108. Caspase-3 (8G10) Rabbit mAb confirms silencing of caspase-3 expression, while the p42 MAPK Antibody is used to control for loading and specificity of caspase-3 siRNA.

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells, untreated (left) or treated with Staurosporine #9953 (1 μM, 3 hr; right), using Cleaved Caspase-9 (Asp330) (E5Z7N) 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 HeLa (human) and NIH/3T3 (mouse) cell extracts, untreated and treated with 1 μM staurosporine (3 hr) in vivo, using Caspase-3 (8G10) Rabbit mAb.


IP

IP

Immunoprecipitation of cleaved caspase-3 from Jurkat cell extracts untreated (control) or treated with etoposide (25uM 5hrs) (apoptotic) using Caspase-3 (8G10) Rabbit mAb, and western probed with the same antibody.

バックグラウンド

Apoptosis is a regulated physiological process leading to cell death. Caspases, a family of cysteine acid proteases, are central regulators of apoptosis. Initiator caspases (including 2, 8, 9, 10 and 12) are closely coupled to proapoptotic signals, which include FasL, TNF-α, and DNA damage. Once activated, these caspases cleave and activate downstream effector caspases (including 3, 6 and 7), which in turn cleave cytoskeletal and nuclear proteins such as PARP, α-fodrin, DFF and lamin A; inducing apoptosis (1,2).

Formation of a death-inducing signaling complex (DISC) around the receptors for death factors, including FasL and TNF-α, is essential for receptor-mediated apoptosis (3). Upon ligand activation, Fas and TNF-R1 associate with death domain (DD) containing adaptor proteins FADD (Fas associated death domain) (4,5) and TRADD (TNF-R1 associated death domain) (6). In addition to a carboxy-terminal DD, FADD contains an amino-terminal death effector domain (DED) that binds to DEDs and activates initiator caspase 8 (FLICE, Mch5, MACH) and caspase 10 (FLICE2, Mch4) (7-12). TRADD does not contain a DED and therefore must associate with FADD in response to TNF-R1 driven apoptosis (13).

Caspase-9 (ICE-LAP6, Mch6) is activated through the mitochondrial-mediated pathway. Cytochrome c released from mitochondria associates with procaspase-9 (47 kDa)/Apaf-1. Apaf-1 mediated activation of caspase-9 involves proteolytic processing resulting in cleavage at Asp315 and producing a p35 subunit. Another cleavage occurs at Asp330 producing a p37 subunit that can amplify the apoptotic response (14-17).

Caspase-2 (Nedd2/ICH-1) is the nuclear apoptotic respondent to cellular genotoxic stress or mitotic catastrophe. The procaspase is cleaved at Asp316, producing a 14 kDa fragment and a 32 kDa prodomain/large subunit. Subsequent processing at Asp152 and Asp330 produces an 18 kDa large subunit and a 12 kDa small fragment (18). Activation occurs upon recruitment to a complex containing a p53-induced death domain protein, PIDD (19). This suggests that caspase-2 can be a nuclear initiator caspase with a requirement for caspase-9 and caspase-3 activation in downstream apoptotic events (20,22). In apoptotic pathways resulting from UV-induced DNA damage, processing of caspase-2 occurs downstream of mitochondrial dysfunction and of caspase-9 and caspase-3 activation, extending a possible role for caspase-2 as a parallel effector caspase (22).

Caspase-3 (CPP-32, Apoptain, Yama, SCA-1) is a critical executioner of apoptosis and caspase-3 cleavage is a key indicator of initiator caspase activation. Caspase-3 is either partially or totally responsible for the proteolytic cleavage of many key proteins including the nuclear enzyme poly (ADP-ribose) polymerase (PARP) (23). Activation of caspase-3 requires proteolytic processing of its inactive zymogen into activated p17 and p12 fragments (24).

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

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