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Signal Transduction Metabolism Drug metabolism

Human NNMT knockout HeLa cell lysate (ab258537)

Price and availability

331 689 ₸

Availability

Order now and get it on Wednesday March 03, 2021

Human NNMT knockout HeLa cell lysate (ab258537)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)

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Overview

  • Product name

    Human NNMT knockout HeLa cell lysate
    See all NNMT kits
  • Product overview


    Knockout cell lysate achieved by CRISPR/Cas9.

  • Parental Cell Line

    HeLa
  • Organism

    Human
  • Mutation description

    Knockout achieved by using CRISPR/Cas9, 16 bp deletion in exon1 and 1 bp insertion in exon1.
  • Passage number

  • Knockout validation

    Sanger Sequencing, Western Blot (WB)
  • Reconstitution notes

    To use as WB control, resuspend the lyophilizate in 50 µL of LDS* Sample Buffer to have a final concentration of 2 mg/ml. For reducing conditions, we recommend a final concentration of 0.1 M DTT.

    *Usage of SDS sample buffer is not recommended with these lyophilized lysates.

  • Notes

    Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version - found here. Please refer to our lysis protocol for further details on how our lysates are prepared.

    User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

    Access thousands of knockout cell lysates, generated from commonly used cancer cell lines.
    See here for more information on knockout cell lysates.

    Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
    It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

    This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.

  • Tested applications

    Suitable for: WBmore details

Properties

  • Storage instructions

    Store at -80°C. Please refer to protocols.
  • Components 1 kit
    ab261270 - Human NNMT knockout HeLa cell lysate 1 x 100µg
    ab255929 - Human wild-type HeLa cell lysate 1 x 100µg
  • Research areas

    • Signal Transduction
    • Metabolism
    • Drug metabolism
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Drug metabolism
  • Cell type

    epithelial
  • Disease

    Adenocarcinoma
  • Gender

    Female
  • STR Analysis

    Amelogenin X D5S818: 11, 12 D13S317: 12, 13.3 D7S820: 8, 12 D16S539: 9, 10 vWA: 16, 18 TH01: 7 TPOX: 8,12 CSF1PO: 9, 10

Target

  • Function

    Catalyzes the N-methylation of nicotinamide and other pyridines to form pyridinium ions. This activity is important for biotransformation of many drugs and xenobiotic compounds.
  • Tissue specificity

    Predominantly expressed in the liver. A lower expression is seen in the kidney, lung, skeletal muscle, placenta and heart. Not detected in the brain or pancreas.
  • Sequence similarities

    Belongs to the NNMT/PNMT/TEMT family.
  • Cellular localization

    Cytoplasm.
  • Target information above from: UniProt accession P40261 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Alternative names

    • EC 2.1.1.1
    • Nicotinamide N methyltransferase
    • Nicotinamide N-methyltransferase
    • NNMT
    • NNMT_HUMAN
    see all

Properties

  • Storage instructions

    Store at -80°C. Please refer to protocols.
  • Components 1 kit
    ab261270 - Human NNMT knockout HeLa cell lysate 1 x 100µg
    ab255929 - Human wild-type HeLa cell lysate 1 x 100µg
  • Research areas

    • Signal Transduction
    • Metabolism
    • Drug metabolism
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Drug metabolism
  • Cell type

    epithelial
  • Disease

    Adenocarcinoma
  • Gender

    Female
  • STR Analysis

    Amelogenin X D5S818: 11, 12 D13S317: 12, 13.3 D7S820: 8, 12 D16S539: 9, 10 vWA: 16, 18 TH01: 7 TPOX: 8,12 CSF1PO: 9, 10

Images

  • Western blot - Human NNMT knockout HeLa cell lysate (ab258537)
    Western blot - Human NNMT knockout HeLa cell lysate (ab258537)

    Lane 1:Wild-type HeLa cell lysate (20 ug)
    Lane 2:NNMT knockout HeLa cell lysate (20 ug)
    Lane 3:A549 cell lysate (20 ug)

    ab119758 was shown to specifically react with NNMT in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265700 (knockout cell lysate ab258537) was used. Wild-type and NNMT knockout samples were subjected to SDS-PAGE. ab119758 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) were incubated at room temperature for 2.5 hours at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Sanger Sequencing - Human NNMT knockout HeLa cell lysate (ab258537)
    Sanger Sequencing - Human NNMT knockout HeLa cell lysate (ab258537)

    Allele-1: 16 bp deletion in exon1

     

  • Sanger Sequencing - Human NNMT knockout HeLa cell lysate (ab258537)
    Sanger Sequencing - Human NNMT knockout HeLa cell lysate (ab258537)

    Allele-2: 1 bp insertion in exon1

     

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

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