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品牌：AdipoGen   货号：AG-20B-0042-C100  

检测炎症小体激活的金标准

独特的抗体应用于WB实验中检测活化的小鼠Caspase-1

Caspase-1是描述炎症的最佳caspase。它能提炼细胞因子interleukin-1β（IL-1β）和L-18，并诱导pyroptotic 细胞死亡。Caspase-1由多蛋白复合物激活，由于在不同的炎症中都能检测到，所以被称为针对多种刺激的炎症小体。NLRC4对胞质鞭毛蛋白反应，鼠源NLRP1b对炭疽致死毒素反应，AlM2对细胞质DNA反应，NLRP3则对包括晶体在内各种激动剂反应。

名称：抗小鼠Caspase-1 （p20）单抗 炎症小体 AG-20B-0042-C100
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品牌货号产品名称规格
AdipogenAG-20B-0042-C100

anti-Caspase-1 (p20) (mouse), mAb (Casper-1)

抗小鼠Caspase-1 （p20）单抗（Casper-1）

100 µg

产品说明书：

AG_20B_0042_Western_Blot_Protocol.pdf

AG-20B-0042-anti-Caspase-1-p20-mouse-mAb-Casper-1.pdf


·特点

·纯化的小鼠单克隆抗体（mAbs）

·识别小鼠caspase-1的内源性全长&激活的p20片段。

·监控炎症小体活性的最好工具

·经过炎症领域专家的测试验证


·案例分析

2314482640303043.jpg

图1：抗体anti-Caspase-1（p20）（mouse），mAb（Casper-1）应用于免疫印迹实验中检测Mouse Caspase-1（p20）。

方法：对细胞提取物和从野生型小鼠分化骨髓来源树突状细胞（BMDCs）进行分化培养，用5μM（Nigericin）处理30分钟，使用WB方法检测细胞上清中NLRP3-/-和caspase-1-/-小鼠是否活化并表达蛋白。在还原条件下，用SDS-PAGE分离细胞提取物和上清液，转移到硝酸纤维素膜后，用anti-Caspase-1（（p20）（mouse），mAb（Casper-1）（1ug/mL）育，最后化学发光进行检测。


2314482640524230.jpg

图2：抗体 anti-Caspase-1（p20）（mouse），mAb（Casper-1）应用于免疫组化实验中染色小鼠脾的内源性小鼠Caspase-1。

方法：通过标准免疫组化（用柠檬酸钠进行抗原修复），使用anti-Caspase-1（p20）（mouse），mAb（Casper-1）（Prod.No.AG-20B-0042）（1：500），使小来源的Caspase-1 KO（左）或者WT（右）的小鼠脾组织（石蜡切片）染色。


Product Details
SynonymsInterleukin-1β Convertase; IL-1BC; Interleukin-1β-converting Enzyme; ICE
Product Type

Monoclonal Antibody


Properties
CloneCasper-1
IsotypeMouse IgG1
Source/HostPurified from concentrated hybridoma tissue culture supernatant.
Immunogen/AntigenRecombinant mouse caspase-1.
Application

Western Blot (see online protocol): (1μg/ml) (no need to precipitate the cell supernatant for the detection of caspase-1 (mouse) upon inflammasome activation)
Immunohistochemistry: (1:500; paraffin sections)
Immunoprecipitation: (1:200)

CrossreactivityMouse
Specificity

Recognizes endogenous full-length and activated (p20 fragment) mouse caspase-1.

Purity≥95% (SDS-PAGE)
Purity DetailProtein G-affinity purified.
Concentration1mg/ml
FormulationLiquid. In PBS containing 10% glycerol and 0.02% sodium azide.
Isotype Negative Control

Mouse IgG1 Isotype Control


Shipping and Handling
ShippingBLUE ICE
Short Term Storage+4°C
Long Term Storage-20°C
Handling AdviceAfter opening, prepare aliquots and store at -20°C.
Avoid freeze/thaw cycles.
Use/Stability

Stable for at least 1 year after receipt when stored at -20°C.


·参考文献

  1. Measuring the inflammasome: O. Gross; Methods Mol. Biol. 844, 199 (2012)
  2. Inflammasome Activators Induce Interleukin-1alpha Secretion via Distinct Pathways with Differential Requirement for the Protease Function of Caspase-1: O. Gross, et al.; Immunity 36, 388 (2012)
  3. Omega-3 Fatty Acids Prevent Inflammation and Metabolic Disorder through Inhibition of NLRP3 Inflammasome Activation: Y. Yan, et al.; Immunity 38, 1154 (2013)
  4. LRRFIP2 negatively regulates NLRP3 inflammasome activation in macrophages by promoting Flightless-I-mediated caspase-1 inhibition: J. Jin, et al.; Nat. Commun. 4, 2075 (2013)
  5. iGLuc: a luciferase-based inflammasome and protease activity reporter: E. Bartok, et al.; Nat. Methods 10, 147 (2013)
  6. Receptor interacting protein kinase 2–mediated mitophagy regulates inflammasome activation during virus infection: C. Lupfer, et al.; Nat. Immunol. 14, 480 (2013)
  7. Activation of the NLRP3 inflammasome by IAV virulence protein PB1-F2 contributes to severe pathophysiology and disease: J.L. McAuley, et al.; PLoS Pathog. 9, e1003392 (2013)
  8. Caspase-1 activity affects AIM2 speck formation/stability through a negative feedback loop: C. Juruj, et al.; Front. Cell. Infect. Microbiol. 3, 14 (2013)
  9. Inflammasome Activation by Altered Proteostasis: J.N. Shin, et al.; J. Biol. Chem. 288, 35886 (2013)
  10. Immunoblotting for active caspase-1: C. Jakobs, et al.; Methods Mol. Biol. 1040, 103 (2013)
  11. Inflammasome activation and inhibition in primary murine bone marrow-derived cells, and assays for IL-1α, IL-1β, and caspase-1: K.s. Schneider, et al.; Methods Mol. Biol. 1040, 117 (2013)
  12. FADD and caspase-8 mediate priming and activation of the canonical and noncanonical Nlrp3 inflammasomes: P. Gurung, et al.; J. Immunol. 192, 1835 (2014)
  13. Salmonella exploits NLRP12-dependent innate immune signaling to suppress host defenses during infection: Md. H. Zaki, et al.; PNAS 111, 385 (2014)
  14. XIAP Restricts TNF- and RIP3-Dependent Cell Death and Inflammasome Activation: M. Yabal, et al.; Cell Rep. 7, 1796 (2014)
  15. The adaptor ASC has extracellular and 'prionoid' activities that propagate inflammation: B.S. Franklin, et al.; Nat. Immunol. 15, 727 (2014)
  16. The NLRP3 inflammasome is released as a particulate danger signal that amplifies the inflammatory response: A. Baroja-Mazo, et al.; Nat. Immunol. 15, 738 (2014)
  17. Caspase-1 activation by NLRP3 inflammasome dampens IL-33-dependent house dust mite-induced allergic lung inflammation: F. Madouri, et al.; J. Mol. Cell Biol. 7, 351 (2015)
  18. ZBP1/DAI is an innate sensor of influenza virus triggering the NLRP3 inflammasome and programmed cell death pathways: T. Kuriakose, et al.; Sci. Immunol. 1, aag2045 (2016)
  19. Assessing Caspase-1 Activation: B. Guey & V. Petrilli; Methods Mol. Biol. 1417, 197 (2016)
  20. IL-1beta and caspase-1 drive autoinflammatory disease independently of IL-1alpha or caspase-8 in a mouse model of familial mediterranean fever: D. Sharma, et al.; Am. J. Pathol. 187, 236 (2017)
  21. The DNA inflammasome in human myeloid cells is initiated by a STING-cell death program upstream of NLRP3: M.M. Gaidt, et al.; Cell 171, 1110 (2017)
  22. Inhibition of Dpp8/9 Activates the Nlrp1b Inflammasome: M.C. Okondo, et al.; Cell Chem. Biol. 25, 1 (2018)
  23. Complement Mediated Activation of the NLRP3 Inflammasome and its Inhibition by AAV Mediated Delivery of CD59 in a Model of Uveitis: B. Kumar, et al.; Mol. Ther. (2018), doi: 10.1016/j.ymthe.2018.03.012
  24. IRF8 Regulates Transcription of Naips for NLRC4 Inflammasome Activation: R. Karki, et al.; Cell 173, 1 (2018)
  25. N-terminal degradation activates the NLRP1B inflammasome: A.J. Chui, et al.; Science 364, 82 (2019)
  26. Functional degradation: A mechanism of NLRP1 inflammasome activation by diverse pathogen enzymes: Science 364, eaau1330 (2019)
  27. NLRP3 inflammasome activation drives tau pathology: C. Ising, et al.; Nature (Epub ahead of Print) (2019)
  28. Caspase-6 Is a Key Regulator of Innate Immunity, Inflammasome Activation, and Host Defense: M. Zheng, et al.; Cell (Epub ahead of print) (2020)


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