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Human ASS1 knockout HeLa cell pellet (ab279021)

Overview

  • Product name

    Human ASS1 knockout HeLa cell pellet
    See all ASS1 kits

  • Product overview

    Abcam’s knockout cell pellets give you access to native proteins, without the need to culture cells. Our knockout cell pellets are prepared from our single-gene knockout cell lines and provide an additional offering to our cell lysates.

    Cells are snap-frozen to provide high quality pellets that are suitable for extraction with alternative lysis buffers or for preparation of lysates from subcellular fractions. Our knockout cell pellets are suitable for a variety of applications, including PCR, gene expression profiling and DNA library preparation.

  • Parental Cell Line

    HeLa

  • Organism

    Human

  • Mutation description

    Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 3 and Insertion of the selection cassette in exon 3.

  • Passage number

  • Knockout validation

    Sanger Sequencing, Western Blot (WB)

  • Notes

    Pellet size: 5 million cells/vial.

    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

Target

  • Pathway

    Amino-acid biosynthesis; L-arginine biosynthesis; L-arginine from L-ornithine and carbamoyl phosphate: step 2/3.
    Nitrogen metabolism; urea cycle; (N(omega)-L-arginino)succinate from L-aspartate and L-citrulline: step 1/1.

  • Involvement in disease

    Defects in ASS1 are the cause of citrullinemia type 1 (CTLN1) [MIM:215700]. Citrullinemia belongs to the urea cycle disorders. It is an autosomal recessive disease characterized primarily by elevated serum and urine citrulline levels. Ammonia intoxication is another manifestation. CTLN1 usually manifests in the first few days of life. Affected infants appear normal at birth, but as ammonia builds up in the body they present symptoms such as lethargy, poor feeding, vomiting, seizures and loss of consciousness. Less commonly, a milder CTLN1 form can develop later in childhood or adulthood.

  • Sequence similarities

    Belongs to the argininosuccinate synthase family. Type 1 subfamily.
  • Information by UniProt

  • Alternative names

    • Argininosuccinate synthase
    • Argininosuccinate synthase 1
    • Argininosuccinate synthetase 1
    • ASS
    • Ass-1
    • ass1
    • ASSA
    • ASSY_HUMAN
    • Citrulline aspartate ligase
    • Citrulline--aspartate ligase
    • CTLN1
    see all

Properties

Images

  • Western blot - Human ASS1 knockout HeLa cell pellet (ab279021)
    Western blot - Human ASS1 knockout HeLa cell pellet (ab279021)
    Lane 1: Wild-type HeLa cell lysate (20µg)

    Lane 2: ASS1 knockout HeLa cell lysate (20µg)
    Lanes 1- 2: Merged signal (red and green). Green - ab170952 observed at 47 kDa. Red - loading control ab8245 observed at 37 kDa.
    ab170952 Anti-ASS1 antibody [EPR12398] was shown to specifically react with ASS1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab264989 (knockout cell lysate ab257143) was used. Wild-type and ASS1 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab170952 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
  • Western blot - Human ASS1 knockout HeLa cell pellet (ab279021)
    Western blot - Human ASS1 knockout HeLa cell pellet (ab279021)
    Lane 1: Wild-type HeLa cell lysate (20µg)

    Lane 2: ASS1 knockout HeLa cell lysate (20µg)
    Lanes 1- 2: Merged signal (red and green). Green - ab170900 observed at 47 kDa. Red - loading control ab8245 observed at 37 kDa.
    ab170900 Anti-ASS1 antibody [EPR12399(B)] - C-terminal was shown to specifically react with ASS1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab264989 (knockout cell lysate ab257143) was used. Wild-type and ASS1 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab170900 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
  • Sanger Sequencing - Human ASS1 knockout HeLa cell pellet (ab279021)
    Sanger Sequencing - Human ASS1 knockout HeLa cell pellet (ab279021)

    Allele-1: 1 bp insertion in exon 3

     

  • Sanger Sequencing - Human ASS1 knockout HeLa cell pellet (ab279021)
    Sanger Sequencing - Human ASS1 knockout HeLa cell pellet (ab279021)

    Allele-2: Insertion of the selection cassette in exon 3

     

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