Validated using a knockout cell line

Anti-ASS1 antibody [2B10] (ab124465)

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Western blot - Anti-ASS1 antibody [2B10] (ab124465)
  • Immunocytochemistry - Anti-ASS1 antibody [2B10] (ab124465)
  • Immunocytochemistry/ Immunofluorescence - Anti-ASS1 antibody [2B10] (ab124465)
  • Flow Cytometry - Anti-ASS1 antibody [2B10] (ab124465)
  • ELISA - Anti-ASS1 antibody [2B10] (ab124465)
  • Western blot - Anti-ASS1 antibody [2B10] (ab124465)
  • Western blot - Anti-ASS1 antibody [2B10] (ab124465)

Key features and details

  • Mouse monoclonal [2B10] to ASS1
  • Suitable for: WB, ELISA, Flow Cyt, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Rat, Human, African green monkey
  • Isotype: IgG1

Overview

  • Product name

    Anti-ASS1 antibody [2B10]
    See all ASS1 primary antibodies

  • Description

    Mouse monoclonal [2B10] to ASS1

  • Host species

    Mouse

  • Tested applications

    Suitable for: WB, ELISA, Flow Cyt, ICC/IFmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human, African green monkey

  • Immunogen

    Purified recombinant fragment of Human ASS1, expressed in E. Coli.

  • Positive control

    • ICC: HeLa cells. Flow Cyt: Jurkat cells. WB: Human ASS1 recombinant protein; HAP1, A431, Raji, L1210, MOLT4, Jurkat, A549, NIH 3T3, PC12 and COS7 cell lysates.

  • General notes

    This product was changed from ascites to supernatant. Lot no’s high than GR119162-30 are from Tissue Culture Supernatant

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab124465 in the following tested applications.

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

Application Abreviews Notes
WB
1/500 - 1/2000. Detects a band of approximately 47 kDa (predicted molecular weight: 47 kDa).
ELISA
1/10000.
Flow Cyt
1/200 - 1/400.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
ICC/IF
1/200 - 1/1000.
Notes
WB
1/500 - 1/2000. Detects a band of approximately 47 kDa (predicted molecular weight: 47 kDa).
ELISA
1/10000.
Flow Cyt
1/200 - 1/400.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
ICC/IF
1/200 - 1/1000.

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

  • Database links

    see all

  • Alternative names

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

Images

  • Western blot - Anti-ASS1 antibody [2B10] (ab124465)
    Western blot - Anti-ASS1 antibody [2B10] (ab124465)

    Lane 1: Wild type HAP1 whole cell lysate (20 µg)
    Lane 2: ASS1 knockout HAP1 whole cell lysate (20 µg)
    Lane 3: HeLa whole cell lysate (20 µg)
    Lane 4: Jurkat whole cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab124465 observed at 47 kDa. Red - loading control, ab181602, observed at 37 kDa.

    ab124465 was shown to recognize ASS1 when ASS1 knockout samples were used, along with additional cross-reactive bands. Wild-type and ASS1 knockout samples were subjected to SDS-PAGE. ab124465 and ab181602 (Rabbit anti-GAPDH loading control) were incubated overnight at 4°C at a 1/500 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • Immunocytochemistry - Anti-ASS1 antibody [2B10] (ab124465)
    Immunocytochemistry - Anti-ASS1 antibody [2B10] (ab124465)
    ab124465 staining ASS1 in wild-type HeLa cells (top panel) and ASS1 knockout HeLa cells (ab264989) (bottom panel). The cells were fixed with 100% methanol (5 min) then permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab124465 at 1/1000 dilution and ab6046 (Rabbit polyclonal to beta Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to mouse IgG (Alexa Fluor® 488) (ab150117) at 2 μg/ml (shown in green) and a goat secondary antibody to rabbit IgG (Alexa Fluor® 594) (ab150080) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
    Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).
  • Immunocytochemistry/ Immunofluorescence - Anti-ASS1 antibody [2B10] (ab124465)
    Immunocytochemistry/ Immunofluorescence - Anti-ASS1 antibody [2B10] (ab124465)
    ab124465, at 1/200 dilution, staining ASS1 in HeLa cells by Immunofluorescence (green). Blue: DRAQ5 fluorescent DNA dye.
  • Flow Cytometry - Anti-ASS1 antibody [2B10] (ab124465)
    Flow Cytometry - Anti-ASS1 antibody [2B10] (ab124465)
    ab124465, at 1/200 dilution, staining ASS1 in Jurkat cells (green) by flow cytometry or negative control (red).
  • ELISA - Anti-ASS1 antibody [2B10] (ab124465)
    ELISA - Anti-ASS1 antibody [2B10] (ab124465)
    ELISA using ab124465.
  • Western blot - Anti-ASS1 antibody [2B10] (ab124465)
    Western blot - Anti-ASS1 antibody [2B10] (ab124465)
    All lanes : Anti-ASS1 antibody [2B10] (ab124465) at 1/500 dilution

    Lane 1 : A431 cell lysate
    Lane 2 : Raji cell lysate
    Lane 3 : L1210 cell lysate
    Lane 4 : MOLT4 cell lysate
    Lane 5 : Jurkat cell lysate
    Lane 6 : A549 cell lysate
    Lane 7 : NIH 3T3 cell lysate
    Lane 8 : PC12 cell lysate
    Lane 9 : COS7 cell lysate

    Predicted band size: 47 kDa

  • Western blot - Anti-ASS1 antibody [2B10] (ab124465)
    Western blot - Anti-ASS1 antibody [2B10] (ab124465)
    Anti-ASS1 antibody [2B10] (ab124465) at 1/500 dilution + Human ASS1 recombinant protein (aa: 40-236)

    Predicted band size: 47 kDa

References (8)

Publishing research using ab124465? Please let us know so that we can cite the reference in this datasheet.

ab124465 has been referenced in 8 publications.

  • Soria LR  et al. Beclin-1-mediated activation of autophagy improves proximal and distal urea cycle disorders. EMBO Mol Med 13:e13158 (2021). PubMed: 33369168
  • Kim S  et al. Argininosuccinate synthase 1 suppresses tumor progression through activation of PERK/eIF2a/ATF4/CHOP axis in hepatocellular carcinoma. J Exp Clin Cancer Res 40:127 (2021). PubMed: 33838671
  • Mehazri L  et al. Loss of Fer Jeopardizes Metabolic Plasticity and Mitochondrial Homeostasis in Lung and Breast Carcinoma Cells. Int J Mol Sci 22:N/A (2021). PubMed: 33806191
  • Silberman A  et al. Acid-Induced Downregulation of ASS1 Contributes to the Maintenance of Intracellular pH in Cancer. Cancer Res 79:518-533 (2019). PubMed: 30573518
  • Li T  et al. Oncogenic Kaposi's Sarcoma-Associated Herpesvirus Upregulates Argininosuccinate Synthase 1, a Rate-Limiting Enzyme of the Citrulline-Nitric Oxide Cycle, To Activate the STAT3 Pathway and Promote Growth Transformation. J Virol 93:N/A (2019). PubMed: 30463977
  • Ochocki JD  et al. Arginase 2 Suppresses Renal Carcinoma Progression via Biosynthetic Cofactor Pyridoxal Phosphate Depletion and Increased Polyamine Toxicity. Cell Metab 27:1263-1280.e6 (2018). PubMed: 29754953
  • Lee JS  et al. Urea Cycle Dysregulation Generates Clinically Relevant Genomic and Biochemical Signatures. Cell 174:1559-1570.e22 (2018). PubMed: 30100185
  • Ohshima K  et al. Argininosuccinate Synthase 1-Deficiency Enhances the Cell Sensitivity to Arginine through Decreased DEPTOR Expression in Endometrial Cancer. Sci Rep 7:45504 (2017). Human . PubMed: 28358054

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