Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Leukotriene A4 hydrolase/LTA4H antibody [EPR5713] (ab133512)

Submit a review Q&A (1)References (4)
Western blot - Anti-Leukotriene A4 hydrolase/LTA4H antibody [EPR5713] (ab133512)
  • Flow Cytometry - Anti-Leukotriene A4 hydrolase/LTA4H antibody [EPR5713] (ab133512)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Leukotriene A4 hydrolase/LTA4H antibody [EPR5713] (ab133512)
  • Western blot - Anti-Leukotriene A4 hydrolase/LTA4H antibody [EPR5713] (ab133512)
  • Western blot - Anti-Leukotriene A4 hydrolase/LTA4H antibody [EPR5713] (ab133512)
  • OI-RD Scanning - Anti-Leukotriene A4 hydrolase/LTA4H antibody [EPR5713] (ab133512)
  • Anti-Leukotriene A4 hydrolase/LTA4H antibody [EPR5713] (ab133512)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR5713] to Leukotriene A4 hydrolase/LTA4H
  • Suitable for: WB, IHC-P, Flow Cyt
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-Leukotriene A4 hydrolase/LTA4H antibody [EPR5713]
    See all Leukotriene A4 hydrolase/LTA4H primary antibodies

  • Description

    Rabbit monoclonal [EPR5713] to Leukotriene A4 hydrolase/LTA4H

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, Flow Cytmore details
    Unsuitable for: IP

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide within Human Leukotriene A4 hydrolase/LTA4H aa 550-650. The exact sequence is proprietary.

  • Positive control

    • WB: HeLa, 293T, T47D and A549 cell lysates. IHC-P: Human kidney tissue. Flow Cyt: Jurkat cells.

  • General notes

     

     

     This product was previously labelled as Leukotriene A4 hydrolase

     

    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.

    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

Applications

Our Abpromise guarantee covers the use of ab133512 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 1/10000 - 1/50000. Predicted molecular weight: 69 kDa.
IHC-P 1/250 - 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Flow Cyt 1/100 - 1/1000.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
  • Application notes
    Is unsuitable for IP.

    • Target

    Images

    • Western blot - Anti-Leukotriene A4 hydrolase/LTA4H antibody [EPR5713] (ab133512)
      Western blot - Anti-Leukotriene A4 hydrolase/LTA4H antibody [EPR5713] (ab133512)
      All lanes : Anti-Leukotriene A4 hydrolase/LTA4H antibody [EPR5713] (ab133512) at 1/1000 dilution

      Lane 1 : Wild-type HEK293T cell lysate
      Lane 2 : LTA4H knockout HEK293T cell lysate
      Lane 3 : T-47D cell lysate
      Lane 4 : A549 cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

      Predicted band size: 69 kDa
      Observed band size: 69 kDa



      Lanes 1-4: Merged signal (red and green). Green - ab133512 observed at 69 kDa. Red - loading control ab8245 observed at 36 kDa.

       ab133512 Anti-Leukotriene A4 hydrolase/LTA4H antibody [EPR5713] was shown to specifically react with Leukotriene in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266467 (knockout cell lysate ab258034) was used. Wild-type and Leukotriene knockout samples were subjected to SDS-PAGE. ab133512 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 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.

    • Flow Cytometry - Anti-Leukotriene A4 hydrolase/LTA4H antibody [EPR5713] (ab133512)
      Flow Cytometry - Anti-Leukotriene A4 hydrolase/LTA4H antibody [EPR5713] (ab133512)
      Overlay histogram showing Jurkat cells stained with ab133512 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab133512, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in Jurkat cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Leukotriene A4 hydrolase/LTA4H antibody [EPR5713] (ab133512)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Leukotriene A4 hydrolase/LTA4H antibody [EPR5713] (ab133512)

      Immunohistochemical analysis of paraffin-embedded Human kidney tissue labelling Leukotriene A4 hydrolase/LTA4H with ab133512 at 1/250 dilution.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Western blot - Anti-Leukotriene A4 hydrolase/LTA4H antibody [EPR5713] (ab133512)
      Western blot - Anti-Leukotriene A4 hydrolase/LTA4H antibody [EPR5713] (ab133512)
      All lanes : Anti-Leukotriene A4 hydrolase/LTA4H antibody [EPR5713] (ab133512) at 1/10000 dilution

      Lane 1 : HeLa cell lysate
      Lane 2 : 293T cell lysate
      Lane 3 : T47D cell lysate
      Lane 4 : A549 cell lysate

      Lysates/proteins at 10 µg per lane.

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

      Predicted band size: 69 kDa

    • Western blot - Anti-Leukotriene A4 hydrolase/LTA4H antibody [EPR5713] (ab133512)
      Western blot - Anti-Leukotriene A4 hydrolase/LTA4H antibody [EPR5713] (ab133512)
      All lanes : Anti-Leukotriene A4 hydrolase/LTA4H antibody [EPR5713] (ab133512) at 1/5000 dilution

      Lane 1 : Mouse brain tissue lysate
      Lane 2 : Mouse heart tissue lysate
      Lane 3 : Mouse kidney tissue lysate
      Lane 4 : Mouse spleen tissue lysate
      Lane 5 : Rat brain tissue lysate
      Lane 6 : Rat heart tissue lysate
      Lane 7 : Rat kidney tissue lysate
      Lane 8 : Rat spleen tissue lysate

      Predicted band size: 69 kDa
      Observed band size: 70 kDa
      why is the actual band size different from the predicted?


      Exposure time: 1 minute
    • OI-RD Scanning - Anti-Leukotriene A4 hydrolase/LTA4H antibody [EPR5713] (ab133512)
      OI-RD Scanning - Anti-Leukotriene A4 hydrolase/LTA4H antibody [EPR5713] (ab133512)
      Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD
    • Anti-Leukotriene A4 hydrolase/LTA4H antibody [EPR5713] (ab133512)
      Anti-Leukotriene A4 hydrolase/LTA4H antibody [EPR5713] (ab133512)

    References (4)

    Publishing research using ab133512? Please let us know so that we can cite the reference in this datasheet.

    ab133512 has been referenced in 4 publications.

    • Chan SJ  et al. Early and Sustained Increases in Leukotriene B4 Levels Are Associated with Poor Clinical Outcome in Ischemic Stroke Patients. Neurotherapeutics 17:282-293 (2020). PubMed: 31520306
    • Tsuda S  et al. Novel mechanism of regulation of the 5-lipoxygenase/leukotriene B4 pathway by high-density lipoprotein in macrophages. Sci Rep 7:12989 (2017). PubMed: 29021582
    • Lin HN & O'Connor JP Immunohistochemical localization of key arachidonic acid metabolism enzymes during fracture healing in mice. PLoS One 9:e88423 (2014). IHC-P ; Mouse . PubMed: 24516658
    • Dixit N  et al. Leukotriene B4 activates intracellular calcium and augments human osteoclastogenesis. Arthritis Res Ther 16:496 (2014). ICC/IF ; Human . PubMed: 25443625

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a question

    Question

    Product code: 133512

    Inquiry: Please can you clarify the difference between ab133512 and ab109434. I wish to quantify Leukotriene A4 hydrolase in human leukocytes by western blot. Which of these two products suits this application better.

    Read More
    Answer

    Thank you for contacting us.

    Both antibodies are raised in rabbit but they are from two different clones (ab109434 from clone EPR5712 compared to ab133512 from clone EPR5713).

    There are slight variations between the two products such as the immunogen sequence, which is amino acids 35-60 for ab109434 and amino acids 575-600 for ab133512 of Human Leukotriene A4 hydrolase.

    Furthermore the products are tested in different species and applications. Antibody ab109434 reacts with rat and human but does not react with mouse and is suitable for WB, IP, IHC-P as well as Flow Cyt. Compared to that antibody ab133512 reacts with mouse, rat and human and was tested in WB, IHC-P, ICC/IF and Flow Cyt applications.


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