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SREBP-1 Transcription Factor Assay试剂盒(ab133125)

Submit a review Q&A (2)References (8)
ELISA ab133125 SREBP-1 Transcription Factor Assay Kit
  • ELISA ab133125 SREBP-1 Transcription Factor Assay Kit
  • Functional Studies - SREBP-1 Transcription Factor Assay Kit (ab133125)
  • Functional Studies - SREBP-1 Transcription Factor Assay Kit (ab133125)
  • Functional Studies - SREBP-1 Transcription Factor Assay Kit (ab133125)

Key features and details

  • Assay type: Semi-quantitative
  • Detection method: Colorimetric
  • Platform: Microplate reader
  • Sample type: Cell culture extracts, Nuclear Extracts

概述

  • 产品名称

    SREBP-1 Transcription Factor Assay试剂盒

  • 检测方法

    Colorimetric

  • 样品类型

    Cell culture extracts, Nuclear Extracts

  • 检测类型

    Semi-quantitative

  • 种属反应性

    与反应: Mouse, Rat, Human

  • 产品概述

    Abcam's SREBP-1 Transcription Factor Assay (ab133125) is a non-radioactive, sensitive method for detecting specific transcription factor DNA binding activity in nuclear extracts.

    A 96-well enzyme-linked immunosorbent assay (ELISA) replaces the cumbersome radioactive electrophoretic mobility shift assay (EMSA). A specific double stranded DNA (dsDNA) sequence containing the SREBP response element is immobilized onto the wells of a 96-well plate. SREBP-1 contained in a nuclear extract binds specifically to the SREBP response element. SREBP-1 is detected by addition of a specific primary antibody directed against SREBP-1. A secondary antibody conjugated to HRP is added to provide a sensitive colorimetric readout at 450 nm. The SREBP-1 Transcription Factor Assay detects human, rat, and murine SREBP-1.

  • 说明

    Lipid homeostasis in vertebrate cells is regulated by a family of transcription factors called sterol regulatory elements binding proteins (SREBPs). SREBPs directly activate the expression of over 30 gened involved in teh synthesis and uptake of cholesterol, fatty cids, triglycerides and phopholipids. SREBP-1 acts primarily to activate genes in fatty acid synthesis. Nuclear SREBP-1 upregulates gene expression of a group of a target lipogenic enzymes such as acetyl CoA carboxylase and fatty acid synthase.

  • 平台

    Microplate reader

性能

相关产品

图片

  • ELISA ab133125 SREBP-1 Transcription Factor Assay Kit
    ELISA ab133125 SREBP-1 Transcription Factor Assay Kit

    HepG2 cells were grown in protein free hybridoma medium overnight (Life Technologies), followed by a 30 minutes incubation with the addition of 1% (2-Hydroxypropyl)-β-cyclodextrin (HPBC; Sigma) or with the supplement of liquid cholesterol (Life Technologies). 10 uL of nuclear extracts (ab113474) were tested, corresponding to 2.5e5 cells per well.

  • ELISA ab133125 SREBP-1 Transcription Factor Assay Kit
    ELISA ab133125 SREBP-1 Transcription Factor Assay Kit

    Positive control (10 or 5 uL) with (10 or 2 uL) or without inhibitor (duplicates; +/- SD).

  • Functional Studies - SREBP-1 Transcription Factor Assay Kit (ab133125)
    Functional Studies - SREBP-1 Transcription Factor Assay Kit (ab133125)
    Comparison of Abcam's SREBP-1 Transcription Factor Assay Kit (ab133125) and SREBP-2 Transcription Factor Assay Kit (ab133114) specificity. The results demonstrate little to no cross reactivity of SREBPs in the opposing assay. Cross Reactivity: (+) SREBP-1a, SREBP-1b, and SREBP-1c
  • Functional Studies - SREBP-1 Transcription Factor Assay Kit (ab133125)
    Functional Studies - SREBP-1 Transcription Factor Assay Kit (ab133125)
    Measurement of SREBP-1 activity in HeLa (squares) and HepG2 (dots) cell lysates.
  • Functional Studies - SREBP-1 Transcription Factor Assay Kit (ab133125)
    Functional Studies - SREBP-1 Transcription Factor Assay Kit (ab133125)
    Quantification of recombinant SREBP-1 from E. coli cell lysates.

文献 (8)

发表研究结果有使用 ab133125？请让我们知道，以便我们可以引用本数据表中的参考文章。

ab133125 被引用在 8 文献中.

  • Alshammari GM  et al. Concomitant Sub-Chronic Administration of Small-Size Gold Nanoparticles Aggravates Doxorubicin-Induced Liver Oxidative and Inflammatory Damage, Hyperlipidemia, and Hepatic Steatosis. Molecules 28:N/A (2023). PubMed: 36677854
  • Dowsett L  et al. Asymmetric dimethylarginine positively modulates calcium-sensing receptor signalling to promote lipid accumulation. Cell Signal 107:110676 (2023). PubMed: 37028778
  • Pham DV & Park PH Adiponectin triggers breast cancer cell death via fatty acid metabolic reprogramming. J Exp Clin Cancer Res 41:9 (2022). PubMed: 34986886
  • Yang Y  et al. LncRNA NEAT1 Potentiates SREBP2 Activity to Promote Inflammatory Macrophage Activation and Limit Hantaan Virus Propagation. Front Microbiol 13:849020 (2022). PubMed: 35495674
  • Zhou J  et al. The Role of PPARd Agosnist GW501516 in Rats with Gestational Diabetes Mellitus. Diabetes Metab Syndr Obes 13:2307-2316 (2020). PubMed: 32669864
  • Chen X  et al. LncRNA NEAT1 promotes hepatic lipid accumulation via regulating miR-146a-5p/ROCK1 in nonalcoholic fatty liver disease. Life Sci 235:116829 (2019). PubMed: 31484042
  • Marwarha G  et al. Palmitate-Induced SREBP1 Expression and Activation Underlies the Increased BACE 1 Activity and Amyloid Beta Genesis. Mol Neurobiol N/A:N/A (2018). PubMed: 30569418
  • Salomone F  et al. Silibinin Restores NAD? Levels and Induces the SIRT1/AMPK Pathway in Non-Alcoholic Fatty Liver. Nutrients 9:N/A (2017). PubMed: 28973994

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Question

I was looking at this assay kit and was wondering if the primary antibody recognizes which part of the protein (Protein as whole or only the nuclear signal). Is this antibody monoclonal or polyclonal? Can you give us the DNA sequence that is coated on the plate, as I would like to rationalize my study after I intend to use it. Also, Please let me know the isoform (full length or nuclear signal) of SREBP1 that was expressed in E.coli. Your graphs for HepG2 cells and HeLa cells say cell lysates, does it mean nuclear/whole cell? Is the competitor ds DNA same as the one that is coated on the plate?

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Answer

The antibody supplied in the SREBP-1 Transcription Factor Assay Kit is polyclonal. We have not mapped which portions of SREBP-1 it detects. The positive control supplied is a clarified cell lysate, so we expect it will contain a mixture of SREBP-1 forms. The antibody in this kit will detect the SREBP-1 that is bound to the immobilized DNA, so if the SREBP-1 is not able to bind to the DNA, it will not be detected in this assay.

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Question

1. What DNA sequence is coated on the plate?
2. What DNA sequence is used as the competitor?
3. Are whole cell lysates or nuclear lysates used from HepG2 and HeLa cells?

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Answer

1. The plate is coated with the dsDNA consensus sequence specific for SREBP. The sequence is 5'TCACCTGA-3'.
2. The competitor DNA sequence is 5'TCACCTGA'.
3. The HepG2 and HeLa lysates are whole cell lysates.




I hope this is helpful. Please contact me again if you have any further questions.

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