Caspase 1 Assay试剂盒(Fluorometric) (ab39412)

概述

  • 产品名称
    Caspase 1 Assay试剂盒(Fluorometric)
    参阅全部 Caspase 1 试剂盒
  • 样品类型
    Tissue Extracts, Cell Lysate
  • 检测类型
    Enzyme activity
  • 检测时间
    2h 00m
  • 产品概述

    Caspase 1 Assay Kit (Fluorometric) (ab39412) provides a simple and convenient method for detecting the activity of caspase 1, which recognizes the sequence YVAD. The assay is based on detection of cleavage of substrate YVAD-AFC (AFC: 7-amino-4-trifluoromethyl coumarin). YVAD-AFC emits blue light (Em=400 nm); upon cleavage of the substrate by caspase-1 or related caspases, free AFC emits a yellow-green fluorescence (Em=505 nm), which can be quantified using a fluorometer or a fluorecence microtiter plate reader. Comparison of the fluorescence from a treated sample with an untreated control allows determination of the fold increase in caspase 1 activity.
    Visit our FAQs page for tips and troubleshooting.

  • 说明

    Caspase 1 (ICE, IL-1beta Converting Enzyme) is the prototypical member of the ICE family of proteases/caspases. Caspase 1 was first identified as a novel protease that generates the proinflammatory cytokine, interleukin 1 beta (IL-1 Beta), by cleaving the pro-interleukin after the Asp116 residue. In addition to its role in the activation of proinflammatory cytokines, Caspase 1 also appears to have functions in some, but not all, types of apoptosis in mammalian cells.

     

  • 经测试应用
    适用于: Functional Studiesmore details

性能

应用

Our Abpromise guarantee covers the use of ab39412 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
Functional Studies Use at an assay dependent concentration.

图片

  • Titration of the caspase 1 (ab39901) (background signal subtracted, duplicates; +/- SD).

  • Badding M.A et al investigated the cytotoxicity of Indium-tin oxide (ITO). Previously,ITO had shown to be cytotoxic in cultured cells and pro-inflammatory in pulmonary animal models. ITO is used to make transparent conductive coatings for touch screens and liquid crystal display electronics. RAW cells were plated at 5 x 105 cells/well and treated with SITO, LPS, or Min-U-sil (cells were first primed with LPS). Cells were washed, lysed, and 100 μg of lysates were assayed using Caspase 1 assay kit (ab39412). PBS was used as a control. All conditions were run in duplicate wells and three independent experiments were performed for each time point.

实验方案

文献

This product has been referenced in:
  • Badding MA  et al. Sintered indium-tin oxide particles induce pro-inflammatory responses in vitro, in part through inflammasome activation. PLoS One 10:e0124368 (2015). Functional Studies . Read more (PubMed: 25874458) »
  • Aruna R  et al. Rutin modulates ASC expression in NLRP3 inflammasome: a study in alcohol and cerulein-induced rat model of pancreatitis. Mol Cell Biochem 396:269-80 (2014). Read more (PubMed: 25060908) »

See all 3 Publications for this product

客户评价及客户问答

The answers to your questions are as follows:


For doing the caspase-1 assay the cells are lysed and then treated with the substrate. Thus, cell permabilization is not an issue.
A reducing environment is required for the caspases to re...

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The lysis buffer in the kit does not contain any protease inhibitors. The assay relies on keeping the samples on ice to minimize degradation, until the assay at 37C. You could add protease inhibitors to a portion of you lysates set aside for western bl...

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ab39412 has sensitivity of 0.05 - 0.5 ng.

There is 1% Triton-x-100 in the lysis buffer. For best results we recommend using the cell lysis buffer provided. If absolutely necessary for the virus, you may use the RIPA buffer with this kit, but results may vary.

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Unfortunately we do not have a sensitivity range determined for this kit. Fold-increase in Caspase-1 activity can be determined by comparing the results of induced samples with the level of the untreated control. Please let me know if yo...

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Thank you for your inquiry. This result would depend on a number of factors including the cell type in which the Capase-1 activation is being studied, the levels of Caspase-1 activation after induction of apoptosis, time-course for Caspase-1 act...

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Thank you for your call today. I have heard back from the lab about your questions: 1) For tissues, homogenize 10 mg of tissue in 200 ul of lysis buffer. Spin out debris and add 50 ul of supernatant into a well of the 96-well plate then proceed from...

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Thank you for your enquiry. The colorimetric assay has sensitivity of about 5 ng and fluorometric is about 10-100 fold more sensitive. Fluorometric assay is recommended for tissue samples. I hope this information helps. Please do not hesitate to...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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