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#12742 Procaspase 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 ホモロジー† 検出分子量 アイソタイプ
Caspase-3 (8G10) Rabbit mAb #9665 20 µl WB, IP H, M, R, Mk 17, 19, 35 Rabbit IgG
Caspase-6 Antibody #9762 20 µl WB H, M, R 15, 35 Rabbit
Caspase-7 (D2Q3L) Rabbit mAb #12827 20 µl WB H, M, R Mk 20, 35 Rabbit IgG
Caspase-8 (1C12) Mouse mAb #9746 20 µl WB, IP H 18, 43, 57 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
Lamin A/C (4C11) Mouse mAb #4777 20 µl WB, IP, IHC-P, IF-F, IF-IC, F H, M, R, Mk 74 (Lamin A), 63 (Lamin C) Mouse IgG2a
PARP Antibody #9542 20 µl WB H, M, R, Mk 89, 116 Rabbit
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl WB Goat
Anti-mouse IgG, HRP-linked Antibody #7076 100 µl WB Horse

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

Applications Key: W=Western Blotting, IP=Immunoprecipitation, IHC-P=Immunohistochemistry (Paraffin), IF-F=Immunofluorescence (Frozen), 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

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 NIH/3T3 cells, untreated or staurosporine-treated (1 µM), and Jurkat cells, untreated or etoposide-treated (25 µM), using PARP Antibody.


Western Blotting

Western Blotting

Western blot analysis of extracts from NIH/3T3 cells, untreated or staurosporine-treated (1 µM) and Jurkat cells, untreated or etoposide-treated (25 µM), using Caspase-6 Antibody.

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 various cell lines using Lamin A/C (4C11) Mouse mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from Jurkat and A20 cells, untreated (-) or treated with Etoposide #2200 (25 μM, overnight; +), using Caspase-7 (D2Q3L) Rabbit 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 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.


Western Blotting

Western Blotting

Western blot analysis of extracts from THP-1 cells, untreated or treated with cycloheximide (CHX, 10 μg/ml, overnight) followed by TNF-α #8902 (20 ng/ml, 4 hr), using Lamin A/C (4C11) Mouse mAb.

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.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Lamin A/C (4C11) Mouse 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.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Lamin A/C (4C11) Mouse mAb.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of HeLa cells (green) using Lamin A/C (4C11) Mouse mAb (solid lines) or a concentration matched Mouse (G3A1) mAb IgG Isotype Control #5415 (dashed lines). Anti-mouse IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4408 was used as a secondary antibody.


IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells using Lamin A/C (4C11) Mouse mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).

IF-F

IF-F

Immunofluorescent analysis of normal rat brain using Lamin A/C (4C11) Mouse mAb (green) and MAP2 Antibody #4542 (red).

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 the 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 like PARP, α-fodrin, DFF and lamin A, and induce apoptosis (1,2).

Caspase-8 (FLICE, Mch5, MACH) and Caspase-9 (ICE-LAP6, Mch6) are initiator caspases. CD95 receptor (Fas/APO-1) and tumor necrosis factor receptor 1 (TNFR1) activate caspase-8, leading to the release of the caspase-8 active fragments, p18 and p10 (3-6). Cytochrome c released from the mitochondria associates with procaspase-9 (47 kDa)/Apaf 1. Apaf-1 mediated activation of caspase-9 involves intrinsic proteolytic processing resulting in cleavage at Asp315 and producing a p35 subunit. Another cleavage occurs at Asp330 producing a p37 subunit that can serve to amplify the apoptotic response (7-11).

Caspase-3 (CPP-32, Apoptain, Yama, SCA-1), Caspase-6 (Mch2), and Caspase-7 (CMH-1, Mch3, ICE-LAP3) are effector caspases (12-16). Activation of caspase-3 requires proteolytic processing of its inactive zymogen/proform into activated p17 and p12 subunits (17). Procaspase-7 is activated through proteolytic processing by upstream caspases at Asp23, Asp198, and Asp206 to produce the mature subunits (14,16). Procaspase-6 is cleaved by caspase-3 at Asp23, Asp179 and Asp193 to form active large (p18) and small (p11) subunits (7).

PARP, a 116 kDa nuclear poly (ADP-ribose) polymerase, appears to be involved in DNA repair in response to environmental stress (18). This protein can be cleaved by many ICE-like caspases in vitro (2,19) and is one of the main cleavage targets of caspase-3 in vivo (17,20). In human PARP, the cleavage occurs between Asp214 and Gly215, which separates the PARP amino-terminal DNA binding domain (24 kDa) from the carboxy-terminal catalytic domain (89 kDa) (17,19). PARP helps cells to maintain their viability; cleavage of PARP facilitates cellular disassembly and serves as a marker of cells undergoing apoptosis (21).

Lamins are nuclear membrane structural components that are important in maintaining normal cell functions such as cell cycle control, DNA replication, and chromatin organization (22-24). Lamin A/C is cleaved by caspase-6 and serves as a marker for caspase-6 activation. During apoptosis, lamin A/C is specifically cleaved into large (41-50 kDa) and small (28 kDa) fragments (24,25). The cleavage of lamins results in nuclear disregulation and cell death (26,27).

使用例
製品をご使用いただいて研究を発表されましたら、ぜひお知らせください

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。

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