胱天蛋白酶Caspase抑制剂 SRB-VAD-FMK(Sulforhodamine B-VAD-FMK) 货号13482-AAT Bioquest荧光染料

上海金畔生物科技有限公司代理AAT Bioquest荧光染料全线产品,欢迎访问AAT Bioquest荧光染料官网了解更多信息。

胱天蛋白酶Caspase抑制剂 SRB-VAD-FMK(Sulforhodamine B-VAD-FMK)

胱天蛋白酶Caspase抑制剂 SRB-VAD-FMK(Sulforhodamine B-VAD-FMK)

胱天蛋白酶Caspase抑制剂 SRB-VAD-FMK(Sulforhodamine B-VAD-FMK)    货号13482 货号 13482 存储条件 在零下15度以下保存, 避免光照
规格 100 Tests 价格 2472
Ex (nm) 559 Em (nm) 577
分子量 874.01 溶剂 DMSO
产品详细介绍

简要概述

胱天蛋白酶Caspase抑制剂 SRB-VAD-FMK(Sulforhodamine B-VAD-FMK)是美国AAT Bioquest生产的用于Caspase的试剂,SRB-VAD是一种红色荧光细胞渗透性胱天蛋白酶抑制剂,可靶向caspase 1,2,3,6,8,9或10.一旦进入细胞内,该抑制剂与活性caspase共价结合,产生绿色荧光。当添加到细胞群中时,SRB-VAD-FMK探针进入每个细胞并共价结合位于活性胱天蛋白酶异二聚体的大亚基上的反应性半胱氨酸残基,从而抑制进一步的酶活性。结合的标记试剂保留在细胞内,而任何未结合的试剂将扩散出细胞并被洗掉。绿色荧光信号是添加试剂时细胞中存在的活性半胱天冬酶量的直接量度。可以通过基于96孔板的荧光测定法,荧光显微术或流式细胞术分析含有结合的标记试剂的细胞。该探针具有与Cy3®和TRITC类似的光谱特性,因此Cy3®和TRITC的滤光片组可以方便地与流式细胞仪或显微镜一起使用。金畔生物是AAT Bioquest的中国代理商,为您提供最优质的胱天蛋白酶Caspase抑制剂 SRB-VAD-FMK(Sulforhodamine B-VAD-FMK)。 

点击查看光谱

产品说明书

样品操作及分析

以下方案仅提供指导,实际情况应根据您的特定需求进行修改。

使用AMC,AFC,pNA,R110和ProRed底物检测胱天蛋白酶的常规方案

1.在DMSO中准备10 mM的储备溶液。

2.如下准备2X半胱天冬酶底物(50 µM)分析溶液:50 µL底物原液,100 µL DTT(1M),400 µL EDTA(100 mM),10 mL Tris Buffer(20 mM),pH = 7.4。

3.将等体积的半胱天冬酶标准品或样品与2X半胱天冬酶底物测定溶液混合,并在室温下孵育至少1小时。

4.使用荧光酶标仪监测荧光,或使用酶标仪监测吸光度。

 

使用细胞渗透性FMK Caspase探针检测细胞Caspase的方案

1.在DMSO中准备2-5 mM的储备溶液。

2.根据需要处理细胞。

3.通过用20 mM Hepes缓冲液(HHBS)在Hanks中稀释DMSO储备溶液(来自步骤2.1),制备2X渗透性半胱天冬酶底物(20 µM)分析溶液。

4.将等体积的处理过的细胞与2X半胱天冬酶底物测定溶液混合(来自步骤2.3),并在37°C,5%CO2的培养箱中孵育细胞至少1小时。

5.用HHBS清洗细胞至少一次。

6.通过流式细胞仪,荧光显微镜或荧光酶标仪监测荧光强度。

 

使用细胞渗透性FMK Caspase探针进行细胞半胱天冬酶测定(仅适用于#13470-13476)

1.通过向小瓶中加入50 µL DMSO制备250X储备液。

2.根据需要处理细胞。

3.以1:250的比例将250 X DMSO储备溶液添加到细胞溶液中(例如2 µL至500 µL细胞),并将细胞在37°C,5%CO2的培养箱中孵育1小时。

4.用HHBS清洗细胞至少一次。

5.通过流式细胞仪,荧光显微镜或荧光酶标仪监测荧光强度。

 

参考文献

Suppression of human T cell proliferation by the caspase inhibitors, z-VAD-FMK and z-IETD-FMK is independent of their caspase inhibition properties
Authors: Lawrence CP, Chow SC.
Journal: Toxicol Appl Pharmacol. (2012)

[Inhibition of elicitation of allergic contact dermatitis by topical use of Z-VAD-FMK, a broad caspase inhibitor: experiment in mice]
Authors: Li YY, Yan CL.
Journal: Zhonghua Yi Xue Za Zhi (2012): 1992

Intracochlear perfusion of leupeptin and z-VAD-FMK: influence of antiapoptotic agents on gunshot-induced hearing loss
Authors: Abaamrane L, Raffin F, Schmerber S, Sendowski I.
Journal: Eur Arch Otorhinolaryngol (2011): 987

Plasmodium falciparum metacaspase PfMCA-1 triggers a z-VAD-fmk inhibitable protease to promote cell death
Authors: Meslin B, Beavogui AH, Fasel N, Picot S.
Journal: PLoS One (2011): e23867

Structure of human caspase-6 in complex with Z-VAD-FMK: New peptide binding mode observed for the non-canonical caspase conformation
Authors: Muller I, Lamers MB, Ritchie AJ, Dominguez C, Munoz-Sanjuan I, Kiselyov A.
Journal: Bioorg Med Chem Lett (2011): 5244

Pretreatment with pancaspase inhibitor (Z-VAD-FMK) delays but does not prevent intraperitoneal heat-killed group B Streptococcus-induced preterm delivery in a pregnant mouse model
Authors: Equils O, Moffatt-Blue C, Ishikawa TO, Simmons CF, Ilievski V, Hirsch E.
Journal: Infect Dis Obstet Gynecol (2009): 749432

[Attenuation of allergic contact dermatitis by Z-VAD-FMK, a broad caspase inhibitor: experiment with mice]
Authors: Li YY, Yan CL, Xu W.
Journal: Zhonghua Yi Xue Za Zhi (2008): 3153

Bodipy-VAD-Fmk, a useful tool to study yeast peptide N-glycanase activity
Authors: Witte MD, Descals CV, de Lavoir SV, Florea BI, van der Marel GA, Overkleeft HS.
Journal: Org Biomol Chem (2007): 3690

Caspase inhibitor Z-VAD-FMK enhances the freeze-thaw survival rate of human embryonic stem cells
Authors: Heng BC, Clement MV, Cao T.
Journal: Biosci Rep (2007): 257

Effects of caspase inhibitors (z-VAD-fmk, z-VDVAD-fmk) on Nile Red fluorescence pattern in 7-ketocholesterol-treated cells: investigation by flow cytometry and spectral imaging microscopy
Authors: Vejux A, Lizard G, Tourneur Y, Riedinger JM, Frouin F, Kahn E.
Journal: Cytometry A (2007): 550

 

相关产品

产品名称 货号
胱天蛋白酶Caspase 3/7荧光底物 FITC-C6-DEVD-FMK 绿 Cat#13406
胱天蛋白酶Caspase抑制剂 FAM-VAD-FMK Cat#13470
胱天蛋白酶Caspase 3/7荧光底物(Z-DEVD)2-R110 绿 Cat#13430

说明书
胱天蛋白酶Caspase抑制剂 SRB-VAD-FMK(Sulforhodamine B-VAD-FMK).pdf

BOC-Asp(OMe)-FMK [Boc-D(OMe)-FMK] CAS 187389-53-3 货号5310-AAT Bioquest荧光染料

上海金畔生物科技有限公司代理AAT Bioquest荧光染料全线产品,欢迎访问AAT Bioquest荧光染料官网了解更多信息。

BOC-Asp(OMe)-FMK [Boc-D(OMe)-FMK] CAS 187389-53-3

BOC-Asp(OMe)-FMK [Boc-D(OMe)-FMK] CAS 187389-53-3

BOC-Asp(OMe)-FMK [Boc-D(OMe)-FMK] CAS 187389-53-3    货号5310 货号 5310 存储条件 在零下15度以下保存, 避免光照
规格 1 mg 价格 1272
Ex (nm) Em (nm)
分子量 263.27 溶剂 DMF
产品详细介绍

简要概述

产品基本信息

货号:5310

产品名称:BOC-Asp(OMe)-FMK [Boc-D(OMe)-FMK]

CAS:187389-53-3

规格:1mg

储存条件:-15℃避光防潮

保质期:12个月

 

产品物理化学光谱特性

分子量:263.27

溶剂:DMF

激发波长(nm):N/A

发射波长(nm):N/A

 

产品介绍

FMK肽是有效的,可透过细胞的,不可逆的capase抑制剂。它可用于检测和监测体外和体内应用中的细胞凋亡。BOC-ASP(OMe)-FMK是开发和制备基于FMK的capase抑制剂和探针的关键中间体。基于Asp(OMe)-FMK的半胱天冬酶抑制剂是细胞可渗透的并且不可逆地结合活化的半胱天冬酶以阻断细胞凋亡。作为分子链锯的胱天蛋白酶是细胞凋亡反应的核心组成部分。半胱天冬酶是半胱氨酸蛋白酶,其天冬氨酸残基在其底物中裂解。14种已知的哺乳动物半胱天冬酶中至少有7种在细胞凋亡中起重要作用。金畔生物是AAT Bioquest的中国代理商,为您提供最优质的BOC-Asp(OMe)-FMK [Boc-D(OMe)-FMK]。

 

参考文献

A glycine-leucine-rich peptide structurally related to the plasticins from skin secretions of the frog Leptodactylus laticeps (Leptodactylidae)
Authors: Conlon JM, Abdel-Wahab YH, Flatt PR, Leprince J, Vaudry H, Jouenne T, Condamine E.
Journal: Peptides (2009): 888

Design of nontoxic analogues of cathelicidin-derived bovine antimicrobial peptide BMAP-27: the role of leucine as well as phenylalanine zipper sequences in determining its toxicity
Authors: Ahmad A, Azmi S, Srivastava RM, Srivastava S, Pandey BK, Saxena R, Bajpai VK, Ghosh JK.
Journal: Biochemistry (2009): 10905

Leucine-rich amelogenin peptide induces osteogenesis by activation of the Wnt pathway
Authors: Warotayanont R, Frenkel B, Snead ML, Zhou Y.
Journal: Biochem Biophys Res Commun (2009): 558

Leucine-rich amelogenin peptide induces osteogenesis in mouse embryonic stem cells
Authors: Warotayanont R, Zhu D, Snead ML, Zhou Y.
Journal: Biochem Biophys Res Commun (2008): 1

Leucine-rich glioma inactivated 3 is involved in amyloid beta peptide uptake by astrocytes and endocytosis itself
Authors: Okabayashi S, Kimura N.
Journal: Neuroreport (2008): 1175

Leucine 7-proline 7 polymorphism in the signal peptide of neuropeptide Y is not a risk factor for exudative age-related macular degeneration
Authors: Kaarniranta K, Holopainen JM, Karvonen MK, Koulu M, Kallio J, Pesonen U, Terasvirta M, Uusitalo H, Immonen I.
Journal: Acta Ophthalmol Scand (2007): 188

Peptide-phospholipid cross-linking reactions: identification of leucine enkephalin-alka(e)nal-glycerophosphatidylcholine adducts by tandem mass spectrometry
Authors: Reis A, Domingues P, Ferrer-Correia AJ, Domingues MR.
Journal: J Am Soc Mass Spectrom (2006): 657

131I-Labeled arginine-arginine-leucine (RRL)-containing cyclic peptide (YCGGRRLGGC) for imaging prostate carcinoma
Authors: Shan L
Journal: (2004)

Leucine-rich amelogenin peptide: a candidate signaling molecule during cementogenesis
Authors: Boabaid F, Gibson CW, Kuehl MA, Berry JE, Snead ML, Nociti FH, Jr., Katchburian E, Somerman MJ.
Journal: J Periodontol (2004): 1126

Cloning the tomato curl3 gene highlights the putative dual role of the leucine-rich repeat receptor kinase tBRI1/SR160 in plant steroid hormone and peptide hormone signaling
Authors: Montoya T, Nomura T, Farrar K, Kaneta T, Yokota T, Bishop GJ.
Journal: Plant Cell (2002): 3163

说明书
BOC-Asp(OMe)-FMK [Boc-D(OMe)-FMK] CAS 187389-53-3.pdf

胱天蛋白酶Caspase抑制剂Z-DEVD-FMK 货号13303-AAT Bioquest荧光染料

上海金畔生物科技有限公司代理AAT Bioquest荧光染料全线产品,欢迎访问AAT Bioquest荧光染料官网了解更多信息。

胱天蛋白酶Caspase抑制剂Z-DEVD-FMK

胱天蛋白酶Caspase抑制剂Z-DEVD-FMK

胱天蛋白酶Caspase抑制剂Z-DEVD-FMK    货号13303 货号 13303 存储条件 在零下15度以下保存, 避免光照
规格 1 mg 价格 1200
Ex (nm) Em (nm)
分子量 668.67 溶剂 DMSO
产品详细介绍

简要概述

产品基本信息

货号:13303

产品名称:胱天蛋白酶Caspase抑制剂Z-DEVD-FMK

规格:1mg

储存条件:-15℃避光防潮

保质期:12个月

 

产品物理化学光谱特性

分子量:668.67

溶剂:DMSO

 

产品介绍

Z-DEVD-FMK是一种不可逆的特异性Caspase-3抑制剂,对caspase-6,caspase-7,caspase-8和caspase-10也具有有效的抑制作用。金畔生物是AAT Bioquest的中国代理商,为您提供最优质的Z-DEVD-FMK。 

 

参考文献

Rescue of Escherichia coli cells from UV-induced death and filamentation by caspase-3 inhibitor.
Authors: Wadhawan, Surbhi and Gautam, Satyendra
Journal: International microbiology : the official journal of the Spanish Society for Microbiology (2019): 369-376

AGEs Induce Apoptosis in Rat Osteoblast Cells by Activating the Caspase-3 Signaling Pathway Under a High-Glucose Environment In Vitro.
Authors: Liu, Jiaqiang and Mao, Jing and Jiang, Yi and Xia, Lunguo and Mao, Lixia and Wu, Yong and Ma, Pan and Fang, Bing
Journal: Applied biochemistry and biotechnology (2016): 1015-27

Cleavage of the calpain inhibitor, calpastatin, during postmortem ageing of beef skeletal muscle.
Authors: Huang, Feng and Huang, Ming and Zhang, Hong and Guo, Bing and Zhang, Dequan and Zhou, Guanghong
Journal: Food chemistry (2014): 1-6

Increased activity of cell surface peptidases in HeLa cells undergoing UV-induced apoptosis is not mediated by caspase 3.
Authors: Piva, Terrence J and Davern, Catherine M and Hall, Paula M and Winterford, Clay M and Ellem, Kay A O
Journal: International journal of molecular sciences (2012): 2650-75

Lysosomal membrane permeabilization is an early event in Sigma-2 receptor ligand mediated cell death in pancreatic cancer.
Authors: Hornick, John R and Vangveravong, Suwanna and Spitzer, Dirk and Abate, Carmen and Berardi, Francesco and Goedegebuure, Peter and Mach, Robert H and Hawkins, William G
Journal: Journal of experimental & clinical cancer research : CR (2012): 41

Caspase 3 blocking avoids the expression of autoantigens triggered by apoptosis in neonatal Balb/c mice skin.
Authors: Ramírez-Sandoval, R and Vazquez-del-Mercado, M and Daneri-Navarro, A and Bollain-y-Goytia, J J and Avalos-Diaz, E and Herrera-Esparza, R
Journal: Reumatismo (2011): 29-37

Disassembly of the JIP1/JNK molecular scaffold by caspase-3-mediated cleavage of JIP1 during apoptosis.
Authors: Vaishnav, Mahesh and MacFarlane, Marion and Dickens, Martin
Journal: Experimental cell research (2011): 1028-39

Role of VPO1, a newly identified heme-containing peroxidase, in ox-LDL induced endothelial cell apoptosis.
Authors: Bai, Yong-Ping and Hu, Chang-Ping and Yuan, Qiong and Peng, Jun and Shi, Rui-Zheng and Yang, Tian-Lun and Cao, Ze-Hong and Li, Yuan-Jian and Cheng, Guangjie and Zhang, Guo-Gang
Journal: Free radical biology & medicine (2011): 1492-500

Evidence for caspase effects on release of cytochrome c and AIF in a model of ischemia in cortical neurons.
Authors: Singh, Maneesh H and Brooke, Sheila M and Zemlyak, Ilona and Sapolsky, Robert M
Journal: Neuroscience letters (2010): 179-83

Proteasome activity profiling: a simple, robust and versatile method revealing subunit-selective inhibitors and cytoplasmic, defense-induced proteasome activities.
Authors: Gu, Christian and Kolodziejek, Izabella and Misas-Villamil, Johana and Shindo, Takayuki and Colby, Tom and Verdoes, Martijn and Richau, Kerstin H and Schmidt, Jürgen and Overkleeft, Hermen S and van der Hoorn, Renier A L
Journal: The Plant journal : for cell and molecular biology (2010): 160-70

说明书
胱天蛋白酶Caspase抑制剂Z-DEVD-FMK.pdf

5-FAM-YVAD-FMK 货号13483-AAT Bioquest荧光染料

上海金畔生物科技有限公司代理AAT Bioquest荧光染料全线产品,欢迎访问AAT Bioquest荧光染料官网了解更多信息。

5-FAM-YVAD-FMK

5-FAM-YVAD-FMK

5-FAM-YVAD-FMK    货号13483 货号 13483 存储条件 在零下15度以下保存, 避免光照
规格 100 Tests 价格 2472
Ex (nm) 493 Em (nm) 517
分子量 854.84 溶剂 DMSO
产品详细介绍

简要概述

5-FAM-YVAD-FMK是美国AAT Bioquest生产的用于Caspase的试剂,FAM-YVAD-FMK,即5-羧基荧光素-Tyr-Val-Ala-Asp-氟甲基酮,在刺激的细胞中不可逆地结合到活性胱天蛋白酶1。 FAM-YVAD-FMK信号的荧光强度与活性半胱天冬酶1的量成正比,可以通过荧光显微镜,流式细胞仪或荧光酶标仪轻松检测。金畔生物是AAT Bioquest的中国代理商,为您提供最优质的5-FAM-YVAD-FMK。 

点击查看光谱

产品说明书

样品操作及分析

以下说明书仅提供指导,实际实验请根据您的特定需求进行修改。

使用AMC,AFC,pNA,R110和ProRed底物的常规胱天蛋白酶测定方案

1.在DMSO中准备10 mM的储备溶液。

2.准备2X半胱天冬酶底物(50 µM)分析溶液:50 µL底物原液,100 µL DTT(1M),400 µL EDTA(100 mM),10 mL Tris Buffer(20 mM),pH = 7.4。

3.将等体积的半胱天冬酶标准品或样品与2X半胱天冬酶底物测定溶液混合,并在室温下孵育至少1小时。

4.使用荧光酶标仪检测荧光或使用酶标仪检测吸光度。

 

使用细胞渗透性FMK Caspase探针进行细胞Caspase测定

1.在DMSO中准备2-5 mM的储备溶液。

2.根据需要处理细胞。

3.通过用20 mM Hepes缓冲液(HHBS)在Hanks中稀释DMSO储备溶液(来自步骤2.1),制备2X渗透性半胱天冬酶底物(20 µM)分析溶液。

4.将等体积的处理过的细胞与2X半胱天冬酶底物测定溶液混合(来自步骤2.3),并在37°C,5%CO2的培养箱中孵育细胞至少1小时。

5.用HHBS清洗细胞至少一次。

6.通过流式细胞仪,荧光显微镜或荧光酶标仪检测荧光强度。

 

使用细胞渗透性FMK Caspase探针进行细胞半胱天冬酶测定(仅适用于#13470-13476)

1.通过向小瓶中加入50 µL DMSO制备250X储备液。

2.根据需要处理细胞。

3.以1:250的比例将250X DMSO储备溶液添加到细胞溶液中(例如2 µL至500 µL细胞),并将细胞在37°C,5%CO2的培养箱中孵育1小时。

4.用HHBS清洗细胞至少一次。

5.通过流式细胞仪,荧光显微镜或荧光酶标仪检测荧光强度。

 

参考文献

The Dietary Flavonoid Fisetin Causes Cell Cycle Arrest, Caspase-Dependent Apoptosis, and Enhanced Cytotoxicity of Chemotherapeutic Drugs in Triple-Negative Breast Cancer Cells
Authors: M. L. Smith
Journal: J Cell Biochem (2016): 1913-25

Contribution of membrane progesterone receptor alpha to the induction of progesterone-mediated apoptosis associated with mitochondrial membrane disruption and caspase cascade activation in Jurkat cell lines
Authors: A. Kon
Journal: Oncol Rep (2013): 1965-70

Inhibition of Caspase-Like Activities Prevents the Appearance of Reactive Oxygen Species and Dark-Induced Apoptosis in the Unicellular Chlorophyte Dunaliella Tertiolecta(1)
Authors: M. Segovia
Journal: J Phycol (2009): 1116-26

Pan-caspase inhibition suppresses polyethylene particle-induced osteolysis
Authors: S. Landgraeber
Journal: Apoptosis (2009): 173-81

TNF induces caspase-dependent inflammation in renal endothelial cells through a Rho- and myosin light chain kinase-dependent mechanism
Authors: X. Wu
Journal: Am J Physiol Renal Physiol (2009): F316-26

Leptin induces interleukin-1beta release from rat microglial cells through a caspase 1 independent mechanism
Authors: E. Pinteaux
Journal: J Neurochem (2007): 826-33

Silvestrol, a potential anticancer rocaglate derivative from Aglaia foveolata, induces apoptosis in LNCaP cells through the mitochondrial/apoptosome pathway without activation of executioner caspase-3 or -7
Authors: S. Kim
Journal: Anticancer Res (2007): 2175-83

Lipopolysaccharide accelerates caspase-independent but cathepsin B-dependent death of human lung epithelial cells
Authors: P. S. Tang
Journal: J Cell Physiol (2006): 457-67

C-terminal region of Bfl-1 induces cell death that accompanies caspase activation when fused with GFP
Authors: W. S. Yang
Journal: J Cell Biochem (2005): 1234-47

Selective caspase activation may contribute to neurological dysfunction after experimental spinal cord trauma
Authors: S. M. Knoblach
Journal: J Neurosci Res (2005): 369-80

 

相关产品

产品名称 货号
5-FAM 5-羧基荧光素 CAS 76823-03-5 Cat#103
6-FAM亚磷酰胺单体 5’-荧光素氨基磷酸酯 Cat#6016
6-FAM 6-羧基荧光素 CAS 3301-79-9 Cat#106

说明书
5-FAM-YVAD-FMK.pdf

胱天蛋白酶Caspase抑制剂 SRB-VAD-FMK(Sulforhodamine B-VAD-FMK) 货号13472-AAT Bioquest荧光染料

上海金畔生物科技有限公司代理AAT Bioquest荧光染料全线产品,欢迎访问AAT Bioquest荧光染料官网了解更多信息。

胱天蛋白酶Caspase抑制剂 SRB-VAD-FMK(Sulforhodamine B-VAD-FMK)

胱天蛋白酶Caspase抑制剂 SRB-VAD-FMK(Sulforhodamine B-VAD-FMK)

胱天蛋白酶Caspase抑制剂 SRB-VAD-FMK(Sulforhodamine B-VAD-FMK)    货号13472 货号 13472 存储条件 在零下15度以下保存, 避免光照
规格 25 Tests 价格 1272
Ex (nm) 559 Em (nm) 577
分子量 874.01 溶剂 DMSO
产品详细介绍

简要概述

胱天蛋白酶Caspase抑制剂 SRB-VAD-FMK(Sulforhodamine B-VAD-FMK)是美国AAT Bioquest生产的用于Caspase的试剂,SRB-VAD是一种红色荧光细胞渗透性胱天蛋白酶抑制剂,可靶向caspase 1,2,3,6,8,9或10.一旦进入细胞内,该抑制剂与活性caspase共价结合,产生绿色荧光。当添加到细胞群中时,SRB-VAD-FMK探针进入每个细胞并共价结合位于活性胱天蛋白酶异二聚体的大亚基上的反应性半胱氨酸残基,从而抑制进一步的酶活性。结合的标记试剂保留在细胞内,而任何未结合的试剂将扩散出细胞并被洗掉。绿色荧光信号是添加试剂时细胞中存在的活性半胱天冬酶量的直接量度。可以通过基于96孔板的荧光测定法,荧光显微术或流式细胞术分析含有结合的标记试剂的细胞。该探针具有与Cy3®和TRITC类似的光谱特性,因此Cy3®和TRITC的滤光片组可以方便地与流式细胞仪或显微镜一起使用。金畔生物是AAT Bioquest的中国代理商,为您提供最优质的胱天蛋白酶Caspase抑制剂 SRB-VAD-FMK(Sulforhodamine B-VAD-FMK)。 

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产品说明书

样品操作及分析

以下方案仅提供指导,实际情况应根据您的特定需求进行修改。

使用AMC,AFC,pNA,R110和ProRed底物检测胱天蛋白酶的常规方案

1.在DMSO中准备10 mM的储备溶液。

2.如下准备2X半胱天冬酶底物(50 µM)分析溶液:50 µL底物原液,100 µL DTT(1M),400 µL EDTA(100 mM),10 mL Tris Buffer(20 mM),pH = 7.4。

3.将等体积的半胱天冬酶标准品或样品与2X半胱天冬酶底物测定溶液混合,并在室温下孵育至少1小时。

4.使用荧光酶标仪监测荧光,或使用酶标仪监测吸光度。

 

使用细胞渗透性FMK Caspase探针检测细胞Caspase的方案

1.在DMSO中准备2-5 mM的储备溶液。

2.根据需要处理细胞。

3.通过用20 mM Hepes缓冲液(HHBS)在Hanks中稀释DMSO储备溶液(来自步骤2.1),制备2X渗透性半胱天冬酶底物(20 µM)分析溶液。

4.将等体积的处理过的细胞与2X半胱天冬酶底物测定溶液混合(来自步骤2.3),并在37°C,5%CO2的培养箱中孵育细胞至少1小时。

5.用HHBS清洗细胞至少一次。

6.通过流式细胞仪,荧光显微镜或荧光酶标仪监测荧光强度。

 

使用细胞渗透性FMK Caspase探针进行细胞半胱天冬酶测定(仅适用于#13470-13476)

1.通过向小瓶中加入50 µL DMSO制备250X储备液。

2.根据需要处理细胞。

3.以1:250的比例将250 X DMSO储备溶液添加到细胞溶液中(例如2 µL至500 µL细胞),并将细胞在37°C,5%CO2的培养箱中孵育1小时。

4.用HHBS清洗细胞至少一次。

5.通过流式细胞仪,荧光显微镜或荧光酶标仪监测荧光强度。

 

参考文献

Suppression of human T cell proliferation by the caspase inhibitors, z-VAD-FMK and z-IETD-FMK is independent of their caspase inhibition properties
Authors: Lawrence CP, Chow SC.
Journal: Toxicol Appl Pharmacol. (2012)

[Inhibition of elicitation of allergic contact dermatitis by topical use of Z-VAD-FMK, a broad caspase inhibitor: experiment in mice]
Authors: Li YY, Yan CL.
Journal: Zhonghua Yi Xue Za Zhi (2012): 1992

Intracochlear perfusion of leupeptin and z-VAD-FMK: influence of antiapoptotic agents on gunshot-induced hearing loss
Authors: Abaamrane L, Raffin F, Schmerber S, Sendowski I.
Journal: Eur Arch Otorhinolaryngol (2011): 987

Plasmodium falciparum metacaspase PfMCA-1 triggers a z-VAD-fmk inhibitable protease to promote cell death
Authors: Meslin B, Beavogui AH, Fasel N, Picot S.
Journal: PLoS One (2011): e23867

Structure of human caspase-6 in complex with Z-VAD-FMK: New peptide binding mode observed for the non-canonical caspase conformation
Authors: Muller I, Lamers MB, Ritchie AJ, Dominguez C, Munoz-Sanjuan I, Kiselyov A.
Journal: Bioorg Med Chem Lett (2011): 5244

Pretreatment with pancaspase inhibitor (Z-VAD-FMK) delays but does not prevent intraperitoneal heat-killed group B Streptococcus-induced preterm delivery in a pregnant mouse model
Authors: Equils O, Moffatt-Blue C, Ishikawa TO, Simmons CF, Ilievski V, Hirsch E.
Journal: Infect Dis Obstet Gynecol (2009): 749432

[Attenuation of allergic contact dermatitis by Z-VAD-FMK, a broad caspase inhibitor: experiment with mice]
Authors: Li YY, Yan CL, Xu W.
Journal: Zhonghua Yi Xue Za Zhi (2008): 3153

Bodipy-VAD-Fmk, a useful tool to study yeast peptide N-glycanase activity
Authors: Witte MD, Descals CV, de Lavoir SV, Florea BI, van der Marel GA, Overkleeft HS.
Journal: Org Biomol Chem (2007): 3690

Caspase inhibitor Z-VAD-FMK enhances the freeze-thaw survival rate of human embryonic stem cells
Authors: Heng BC, Clement MV, Cao T.
Journal: Biosci Rep (2007): 257

Effects of caspase inhibitors (z-VAD-fmk, z-VDVAD-fmk) on Nile Red fluorescence pattern in 7-ketocholesterol-treated cells: investigation by flow cytometry and spectral imaging microscopy
Authors: Vejux A, Lizard G, Tourneur Y, Riedinger JM, Frouin F, Kahn E.
Journal: Cytometry A (2007): 550

 

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说明书
胱天蛋白酶Caspase抑制剂 SRB-VAD-FMK(Sulforhodamine B-VAD-FMK).pdf