Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-DDAH1 antibody [EPR13922] (ab180599)

Reviews (1) Submit a question References (3)
Western blot - Anti-DDAH1 antibody [EPR13922] (ab180599)
  • Western blot - Anti-DDAH1 antibody [EPR13922] (ab180599)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DDAH1 antibody [EPR13922] (ab180599)
  • Flow Cytometry - Anti-DDAH1 antibody [EPR13922] (ab180599)
  • Immunocytochemistry/ Immunofluorescence - Anti-DDAH1 antibody [EPR13922] (ab180599)
  • Immunoprecipitation - Anti-DDAH1 antibody [EPR13922] (ab180599)
  • Anti-DDAH1 antibody [EPR13922] (ab180599)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR13922] to DDAH1
  • Suitable for: Flow Cyt, WB, IHC-P, ICC/IF, IP
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-DDAH1 antibody [EPR13922]
    See all DDAH1 primary antibodies

  • Description

    Rabbit monoclonal [EPR13922] to DDAH1

  • Host species

    Rabbit

  • Tested applications

    Suitable for: Flow Cyt, WB, IHC-P, ICC/IF, IPmore details

  • Species reactivity

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

  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human DDAH1 aa 200 to the C-terminus. The exact sequence is proprietary.
    Database link: O94760

  • Positive control

    • WB: HuVEC, A431 and HepG2 Human fetal brain, liver, kidney or 293 lysate, Human ovarian carcinoma tissue, 293 cells.

  • General notes

     

     

    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
    For more information see here.

    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.

    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

  • 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

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant
  • Concentration information loading...

  • Purity

    Tissue culture supernatant

  • Clonality

    Monoclonal

  • Clone number

    EPR13922

  • Isotype

    IgG

  • Research areas

Applications

Our Abpromise guarantee covers the use of ab180599 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
Flow Cyt 1/20.
WB 1/10000 - 1/50000. Detects a band of approximately 37 kDa.
IHC-P 1/50. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

For antigen retrieval heat up to 98 °C, below boiling, and then let cool for 10-20 min.
ICC/IF 1/100.
IP 1/40.

Target

Images

  • Western blot - Anti-DDAH1 antibody [EPR13922] (ab180599)
    Western blot - Anti-DDAH1 antibody [EPR13922] (ab180599)
    All lanes : Anti-DDAH1 antibody [EPR13922] (ab180599) at 1/10000 dilution

    Lane 1 : A431 wild-type cell lysate
    Lane 2 : A431 DDAH1 knockout cell lysate
    Lane 3 : HuVEC cell lysate
    Lane 4 : HepG2 cell lysate

    Lysates/proteins at 40 µg per lane.

    Performed under reducing conditions.

    Observed band size: 37 kDa
    why is the actual band size different from the predicted?



    Lanes 1 - 4: Merged signal (red and green). Green - ab180599 observed at 37 kDa. Red - loading control, ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.

    ab180599 was shown to react with DDAH1 in A431 wild-type cells in Western blot. Loss of signal was observed when DDAH1 knockout sample was used. A431 wild-type and DDAH1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% Milk in TBS-T (0.1% Tween®) before incubation with ab180599 and ab7291 (Mouse anti-Alpha Tubulin [DM1A] overnight at 4°C at a 1 in 10000 dilution and a 1 in 20000 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 in 20000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-DDAH1 antibody [EPR13922] (ab180599)
    Western blot - Anti-DDAH1 antibody [EPR13922] (ab180599)
    All lanes : Anti-DDAH1 antibody [EPR13922] (ab180599) at 1/20000 dilution

    Lane 1 : Human fetal brain lysate at 20 µg
    Lane 2 : Human fetal liver lysate at 20 µg
    Lane 3 : Human fetal kidney lysate at 20 µg
    Lane 4 : 293 lysate at 10 µg

    Secondary
    All lanes : Goat Anti-Rabbit IgG (H+L) Peroxidase conjugated at 1/1000 dilution

    Observed band size: 37 kDa why is the actual band size different from the predicted?

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DDAH1 antibody [EPR13922] (ab180599)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DDAH1 antibody [EPR13922] (ab180599)

    Immunohistochemical analysis of paraffin embedded Human ovarian carcinoma tissue labeling DDAH1 with ab180599 at a 1/50 dilution. Prediluted HRP Polymer for Rabbit IgG secondary used. Counterstained with Hematoxylin.

    Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

  • Flow Cytometry - Anti-DDAH1 antibody [EPR13922] (ab180599)
    Flow Cytometry - Anti-DDAH1 antibody [EPR13922] (ab180599)

    ab180599 staining DDAH1 in the human cell line 293 (human embryonic kidney) by flow cytometry. Cells were fixed with 4% paraformaldehyde, permabilised with 90% methanol and the sample was incubated with the primary antibody at a dilution of 1/20. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/2000 was used as the secondary antibody.

    Isoytype control: Rabbit monoclonal IgG (Black)

    Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)

  • Immunocytochemistry/ Immunofluorescence - Anti-DDAH1 antibody [EPR13922] (ab180599)
    Immunocytochemistry/ Immunofluorescence - Anti-DDAH1 antibody [EPR13922] (ab180599)

    Immunofluorescence analysis of 293 cells (fixative 4% paraformaldehyde) labeling DDAH1 with ab180599 at a 1/100 dilution (left image), and counterstained with Dapi (right image). Goat anti rabbit IgG (Dylight 555) secondary used at a 1/200 diution.

  • Immunoprecipitation - Anti-DDAH1 antibody [EPR13922] (ab180599)
    Immunoprecipitation - Anti-DDAH1 antibody [EPR13922] (ab180599)

    Immunoprecipitation analysis of Human fetal kidney lysate labeling DDAH1 with ab180599 at a 1/40 dilution. Goat Anti-Rabbit IgG (H+L) Peroxidase conjugated secondary used at a 1/1000 dilution.

  • Anti-DDAH1 antibody [EPR13922] (ab180599)
    Anti-DDAH1 antibody [EPR13922] (ab180599)

References (3)

Publishing research using ab180599? Please let us know so that we can cite the reference in this datasheet.

ab180599 has been referenced in 3 publications.

  • Yang D  et al. Dihydromyricetin increases endothelial nitric oxide production and inhibits atherosclerosis through microRNA-21 in apolipoprotein E-deficient mice. J Cell Mol Med 24:5911-5925 (2020). PubMed: 32301289
  • Tong C  et al. Shock waves increase pulmonary vascular leakage, inflammation, oxidative stress, and apoptosis in a mouse model. Exp Biol Med (Maywood) 243:934-944 (2018). PubMed: 29984607
  • Wang H  et al. DDAH1 plays dual roles in PM2.5 induced cell death in A549 cells. Biochim Biophys Acta 1860:2793-801 (2016). PubMed: 26996393

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-DDAH1 antibody [EPR13922]

 Good
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Kidney)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM Citrate pH6.0
Permeabilization
No
Specification
Kidney
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative
Paraformaldehyde
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Submitted Dec 22 2016

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