Recombinant Anti-ASS1 antibody [EPR12398] - BSA and Azide free (ab231684)
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 - BSA and Azide free
- Suitable for: WB, ICC/IF, IP, Flow Cyt, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Human
Overview
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Product name
Anti-ASS1 antibody [EPR12398] - BSA and Azide free
See all ASS1 primary antibodies -
Description
Rabbit monoclonal [EPR12398] to ASS1 - BSA and Azide free -
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
Synthetic peptide within Human ASS1 aa 1-100 (Cysteine residue). The exact sequence is proprietary.
Database link: P00966 -
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: HeLa cells. IP: HeLa cells.
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General notes
ab231684 is the carrier-free version of ab170952. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.
Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
ab231684 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.
Maxpar® is a trademark of Fluidigm Canada Inc.
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.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
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, as well as customer reviews and Q&As.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: 100% PBS -
Carrier free
Yes -
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
Our Abpromise guarantee covers the use of ab231684 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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WB | Use at an assay dependent concentration. Predicted molecular weight: 47 kDa. | |
ICC/IF | Use at an assay dependent concentration. | |
IP | Use at an assay dependent concentration. | |
Flow Cyt | Use at an assay dependent concentration. | |
IHC-P | Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
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 kDaThis data was developed using the same antibody clone in a different buffer formulation (ab170952).
Lanes 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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This data was developed using the same antibody clone in a different buffer formulation (ab170952). 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 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab170952).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ASS1 antibody [EPR12398] - BSA and Azide free (ab231684)
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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab170952).
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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.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab170952).
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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).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab170952).
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Immunocytochemistry/ Immunofluorescence - Anti-ASS1 antibody [EPR12398] - BSA and Azide free (ab231684)
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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab170952).
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Secondary antibody used is HRP-conjugated anti-rabbit IgG preferentially detecting the non-reduced form of rabbit IgG.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab170952).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ASS1 antibody [EPR12398] - BSA and Azide free (ab231684)
Immunohistochemical (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.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab170952).
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
Certificate of Compliance
References (0)
ab231684 has not yet been referenced specifically in any publications.