Overview

  • Product name

    Anti-ASS1 antibody [EPR12398]
    See all ASS1 primary antibodies
  • Description

    Rabbit monoclonal [EPR12398] to ASS1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ICC/IF, IP, Flow Cyt, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
    Predicted to work with: Cow
  • Immunogen

    within Human ASS1 aa 1-100 (Cysteine residue). The exact sequence is proprietary.
    Database link: P00966

  • Positive control

    • HeLa whole cell lysate (ab150035); Human fetal kidney and Human fetal liver tissue lysates; Human kidney tissue; HeLa cells; Mouse kidney; MCF-7.
  • General notes

    The rat recommendation is based on the WB results. This antibody may not be suitable for IHC with rat samples.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab170952 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 1/20000. Predicted molecular weight: 47 kDa.

For unpurified use at 1/1000 - 1/10000. 

ICC/IF 1/50 - 1/100.
IP 1/10 - 1/100.
Flow Cyt 1/100.

For unpurified use at 1/500 - 1/1000.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IHC-P 1/4000.

See IHC antigen retrieval protocols.

For unpurified use at 1/250 - 1/500. 

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

  • 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

  • All lanes : Anti-ASS1 antibody [EPR12398] (ab170952) at 1/20000 dilution

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : ASS1 knockout HAP1 whole cell lysate
    Lane 3 : HepG2 whole cell lysate
    Lane 4 : HeLa whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 47 kDa
    Observed band size: 47 kDa



    Lanes 1 - 4: Merged signal (red and green). Green - ab170952 observed at 47 kDa. Red - loading control, ab9484, observed at 37 kDa.

    ab170952 was shown to recognize ASS1 in wild-type HAP1 cells as signal was lost at the expected MW in ASS1 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and ASS1 knockout samples were subjected to SDS-PAGE. Ab170952 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/20000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • Immunocytochemistry/ Immunofluorescence analysis of MCF7 (Human breast adenocarcinoma cell line) cells labeling ASS1 with Purified ab170952 at 1:100 dilution (10.2μg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 μg/ml). ab150077 Goat anti rabbit IgG(Alexa Fluor® 488) was used as the secondary antibody at 1:1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

  • ab170952 showing +ve staining in Mouse kidney tissue.

  • All lanes : Anti-ASS1 antibody [EPR12398] (ab170952) at 0.05 µg/ml (Purified)

    Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
    Lane 2 : Human fetal liver lysate
    Lane 3 : Mouse liver lysate
    Lane 4 : Rat liver lysate
    Lane 5 : Mouse kidney lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 47 kDa
    Observed band size: 47 kDa



    Blocking and diluting buffer: 5% NFDM/TBST

  • Flow Cytometry analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling ASS1 with purified ab170952 at 1:100 dilution (10 ug/ml) (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody was used at 1:2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

  • ab170952 (purified) at 1:60 dilution (5ug) immunoprecipitating ASS1 in HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate.
    Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10ug
    Lane 2 (+): ab170952 & HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab170952 in HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
    For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.
    Blocking and diluting buffer: 5% NFDM/TBST.

  • Immunofluorescent analysis of HeLa cells labeling ASS1 using ab170952 at 1/50 dilution (red). DAPI nuclear staining (blue).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse kidney tissue sections labeling ASS1 with Purified ab170952 at 1:4000 dilution (0.25 μg/ml). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using citrate Buffer, PH6. Tissue was counterstained with Hematoxylin. ab97051 Goat Anti-Rabbit IgG H&L (HRP)
    secondary antibody was used at 1:500 dilution. PBS instead of the primary antibody was used as the negative control.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human kidney tissue sections labeling ASS1 with Purified ab170952 at 1:4000 dilution (0.25 μg/ml). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using citrate Buffer, PH6. Tissue was counterstained with Hematoxylin. ab97051 Goat Anti-Rabbit IgG H&L (HRP) secondary antibody was used at 1:500 dilution. PBS instead of the primary antibody was used as the negative control.

  • ab170952 showing +ve staining in Human normal ureter tissue.

  • Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling ASS1 with ab170952 at 1/250 dilution.

  • ab170952 showing -ve staining in Human cervical carcinoma tissue.
  • ab170952 showing -ve staining in Human normal uterus tissue.
  • ab170952 showing -ve staining in Human normal pancreas tissue.
  • Secondary antibody used is HRP-conjugated anti-rabbit IgG preferentially detecting the non-reduced form of rabbit IgG.



    All lanes : Anti-ASS1 antibody [EPR12398] (ab170952) at 1/10 dilution

    Lane 1 : Human fetal liver tissue lysate at 10 µg
    Lane 2 : PBS
  • Flow cytometric analysis of permeabilized Hela cells labeling ASS1 using ab170952 at 1/500 dilution  (red) or a rabbit IgG negative (green).

  • All lanes : Anti-ASS1 antibody [EPR12398] (ab170952) at 1/1000 dilution

    Lane 1 : HeLa cell lysate
    Lane 2 : Fetal liver tissue lysate
    Lane 3 : Fetal kidney tissue lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat anti-rabbit HRP at 1/2000 dilution

    Predicted band size: 47 kDa

References

This product has been referenced in:

See all 2 Publications for this product

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