Recombinant Anti-ASS1 antibody [EPR12398] (ab170952)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR12398] to ASS1
- Suitable for: Flow Cyt (Intra), WB, ICC/IF, IP, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-ASS1 antibody [EPR12398]
See all ASS1 primary antibodies -
Description
Rabbit monoclonal [EPR12398] to ASS1 -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, ICC/IF, IP, IHC-Pmore details -
Species reactivity
Reacts with: Mouse, Rat, Human
Predicted to work with: Cow -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HAP1, HeLa, HepG2 cell lysates. Human fetal kidney and liver tissue lysates. Mouse liver and kidney lysates. Rat liver lysate. ICC/IF: MCF7 and HeLa cells. IHC-P: Human kidney, ureter tissue. Mouse kidney tissue. Flow Cyt (intra): HeLa cells. IP: HeLa cells.
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General notes
The rat recommendation is based on the WB results. This antibody may not be suitable for IHC with rat samples.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR12398 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
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 |
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Flow Cyt (Intra) |
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. |
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WB |
1/20000. Predicted molecular weight: 47 kDa.
For unpurified use at 1/1000 - 1/10000. |
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ICC/IF |
1/50 - 1/100.
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IP |
1/10 - 1/100.
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IHC-P | (1) |
1/4000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
See IHC antigen retrieval protocols. For unpurified use at 1/250 - 1/500. |
Notes |
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Flow Cyt (Intra)
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. |
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. |
IHC-P
1/4000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. See IHC antigen retrieval protocols. For unpurified use at 1/250 - 1/500. |
Target
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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
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Database links
- Entrez Gene: 280726 Cow
- Entrez Gene: 445 Human
- Entrez Gene: 11898 Mouse
- Entrez Gene: 25698 Rat
- Omim: 603470 Human
- SwissProt: P14568 Cow
- SwissProt: P00966 Human
- SwissProt: P16460 Mouse
see all -
Alternative names
- Argininosuccinate synthase 1 antibody
- Argininosuccinate synthase antibody
- Argininosuccinate synthetase 1 antibody
see all
Images
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All lanes : Anti-ASS1 antibody [EPR12398] (ab170952) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : ASS1 knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 47 kDa
Observed band size: 47 kDaLanes 1- 2: Merged signal (red and green). Green - ab170952 observed at 47 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab170952 was shown to 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 HeLa and ASS1 knockout HeLa cell lysates 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) overnight at 4°C at a 1 in 1000 dilution and a 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.
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ab170952 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 ab170952 at 1/100 dilution and ab7291 (Mouse monoclonal to alpha 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 rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (ab150120) 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). -
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 kDaLanes 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.
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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 kDaBlocking and diluting buffer: 5% NFDM/TBST
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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. -
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.
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ab170952 showing +ve staining in Mouse kidney tissue.
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Intracellular 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).
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Immunofluorescent analysis of HeLa cells labeling ASS1 using ab170952 at 1/50 dilution (red). DAPI nuclear staining (blue).
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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 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 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.
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ab170952 showing +ve staining in Human normal ureter tissue.
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Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling ASS1 with ab170952 at 1/250 dilution.
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ab170952 showing -ve staining in Human cervical carcinoma tissue.
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ab170952 showing -ve staining in Human normal uterus tissue.
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ab170952 showing -ve staining in Human normal pancreas tissue.
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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 -
Intracellular flow cytometric analysis of permeabilized Hela cells labeling ASS1 using ab170952 at 1/500 dilution (red) or a rabbit IgG negative (green).
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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
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (8)
ab170952 has been referenced in 8 publications.
- Miao YQ et al. N(6)-adenosine-methyltransferase-14 promotes glioma tumorigenesis by repressing argininosuccinate synthase 1 expression in an m6A-dependent manner. Bioengineered 13:1858-1871 (2022). PubMed: 35012429
- Zhang W et al. The related mechanism of complete Freund's adjuvant-induced chronic inflammation pain based on metabolomics analysis. Biomed Chromatogr 35:e5020 (2021). PubMed: 33159321
- Wang W et al. Tandem Mass Tag-Based Proteomic Analysis of Potential Biomarkers for Hepatocellular Carcinoma Differentiation. Onco Targets Ther 14:1007-1020 (2021). PubMed: 33603407
- Crump NT et al. Chromatin accessibility governs the differential response of cancer and T cells to arginine starvation. Cell Rep 35:109101 (2021). PubMed: 33979616
- Chow JPH et al. A modified arginine-depleting enzyme NEI-01 inhibits growth of pancreatic cancer cells. PLoS One 15:e0231633 (2020). PubMed: 32353864
- Lercher A et al. Type I Interferon Signaling Disrupts the Hepatic Urea Cycle and Alters Systemic Metabolism to Suppress T Cell Function. Immunity 51:1074-1087.e9 (2019). PubMed: 31784108
- Poillet-Perez L et al. Autophagy maintains tumour growth through circulating arginine. Nature 563:569-573 (2018). PubMed: 30429607
- Rabinovich S et al. Diversion of aspartate in ASS1-deficient tumours fosters de novo pyrimidine synthesis. Nature 527:379-383 (2015). ICC/IF . PubMed: 26560030