Validated using a knockout cell line

Anti-Lipoprotein lipase antibody [5D2] (ab93898)

Reviews (2)Q&A (2)References (2)

Key features and details

  • Mouse monoclonal [5D2] to Lipoprotein lipase
  • Suitable for: ICC/IF, Flow Cyt
  • Knockout validated
  • Reacts with: Rat, Cat, Human, Syrian hamster
  • Isotype: IgG1

Overview

  • Product name

    Anti-Lipoprotein lipase antibody [5D2]
    See all Lipoprotein lipase primary antibodies

  • Description

    Mouse monoclonal [5D2] to Lipoprotein lipase

  • Host species

    Mouse

  • Specificity

    This antibody does not recognise hepatic lipase.

  • Tested applications

    Suitable for: ICC/IF, Flow Cytmore details

  • Species reactivity

    Reacts with: Rat, Cat, Human, Syrian hamster

  • Immunogen

    Purified bovine milk lipoprotein lipase protein.

  • Positive control

    • ICC: HeLa cells and mouse cortical glia. Flow Cyt: HeLa cells.

  • General notes

    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 ab93898 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
ICC/IF Use a concentration of 2 - 5 µg/ml.
Flow Cyt Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 

Target

  • Function

    The primary function of this lipase is the hydrolysis of triglycerides of circulating chylomicrons and very low density lipoproteins (VLDL). Binding to heparin sulfate proteogylcans at the cell surface is vital to the function. The apolipoprotein, APOC2, acts as a coactivator of LPL activity in the presence of lipids on the luminal surface of vascular endothelium.

  • Involvement in disease

    Defects in LPL are the cause of lipoprotein lipase deficiency (LPL deficiency) [MIM:238600]; also known as familial chylomicronemia or hyperlipoproteinemia type I. LPL deficiency chylomicronemia is a recessive disorder usually manifesting in childhood. On a normal diet, patients often present with abdominal pain, hepatosplenomegaly, lipemia retinalis, eruptive xanthomata, and massive hypertriglyceridemia, sometimes complicated with acute pancreatitis.

  • Sequence similarities

    Belongs to the AB hydrolase superfamily. Lipase family.
    Contains 1 PLAT domain.

  • Post-translational
    modifications

    Tyrosine nitration after lipopolysaccharide (LPS) challenge down-regulates the lipase activity.

  • Cellular localization

    Cell membrane. Secreted. Locates to the plasma membrane of microvilli of hepatocytes with triacyl-glycerol-rich lipoproteins (TRL). Some of the bound LPL is then internalized and located inside non-coated endocytic vesicles.
  • Information by UniProt

  • Database links

    • Alternative names

      • EC 3.1.1 antibody
      • EC 3.1.1.34 antibody
      • HDLCQ11 antibody
      • LIPD antibody
      • LIPL_HUMAN antibody
      • Lipoprotein lipase antibody
      • LPL antibody
      • LPL protein antibody
      • MGC137861 antibody
      see all

    Images

    • Immunocytochemistry - Anti-Lipoprotein lipase antibody [5D2] (ab93898)
      Immunocytochemistry - Anti-Lipoprotein lipase antibody [5D2] (ab93898)
      ab93898 staining Lipoprotein lipase in wild-type HeLa cells (top panel) and LPL knockout HeLa cells (bottom panel). The cells were fixed with 4% paraformaldehyde (10 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 ab93898 at 5μg/ml concentration and ab6046 (Rabbit polyclonal to beta 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 mouse IgG (Alexa Fluor® 488) (ab150117) at 2 μg/ml (shown in green) and a goat secondary antibody to rabbit IgG (Alexa Fluor® 594) (ab150080) 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).
    • Flow Cytometry - Anti-Lipoprotein lipase antibody [5D2] (ab93898)
      Flow Cytometry - Anti-Lipoprotein lipase antibody [5D2] (ab93898)
      Overlay histogram showing HeLa cells stained with ab93898 (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 (ab93898, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

    • Immunocytochemistry/ Immunofluorescence - Anti-Lipoprotein lipase antibody [5D2] (ab93898)
      Immunocytochemistry/ Immunofluorescence - Anti-Lipoprotein lipase antibody [5D2] (ab93898)

      Immunocytochemistry/Immunofluorescence analysis of mouse cortical glia labelling Lipoprotein lipase (green) with ab93898 at 2µg/ml. Cells were permemabilized with PBS + 0.1% Triton X-100 and blocked with PBS + 10% FBS. Nuclei stained with Hoechst (blue).

    References (2)

    Publishing research using ab93898? Please let us know so that we can cite the reference in this datasheet.

    ab93898 has been referenced in 2 publications.

    • Graham BB  et al. Severe pulmonary hypertension is associated with altered right ventricle metabolic substrate uptake. Am J Physiol Lung Cell Mol Physiol 309:L435-40 (2015). WB, IHC-P ; Rat . PubMed: 26115672
    • Chang SF  et al. Detailed characterization of the binding site of the lipoprotein lipase-specific monoclonal antibody 5D2. J Lipid Res 39:2350-9 (1998). PubMed: 9831623

    Customer reviews and Q&As

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    1-4 of 4 Abreviews or Q&A

    Immunohistochemistry (PFA perfusion fixed frozen sections) abreview for Anti-Lipoprotein lipase antibody [5D2]

     Good
    Abreviews
    abreview image
    Application
    Immunohistochemistry (PFA perfusion fixed frozen sections)
    Sample
    Cat Tissue sections (Liver)
    Antigen retrieval step
    None
    Permeabilization
    Yes - 0.2%TritonX-100 + 0.05%Tween20
    Specification
    Liver
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 23°C
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Feb 05 2020

    Immunohistochemistry (PFA perfusion fixed frozen sections) abreview for Anti-Lipoprotein lipase antibody [5D2]

     Average
    Abreviews
    abreview image
    Application
    Immunohistochemistry (PFA perfusion fixed frozen sections)
    Sample
    Mouse Tissue sections (Heart)
    Antigen retrieval step
    None
    Permeabilization
    Yes - 0.2%TritonX-100 + 0.05%Tween20
    Specification
    Heart
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 23°C
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Feb 05 2020

    Question

    High background in Western blot.

    Read More
    Answer

    Thank you for your call earlier this week and for your patience while I have been in touch with the lab.

    Ab93898 has been cited for use in Western blot in the literature, but we don't have an in-house protocol.I've attached an article that describes the protocol used (see page 2352). It looks like the authorsalso used 1 ug/mL concentration so I think that is fine for you to continue using. Their secondary antibody was diluted 1:50,000, and the optimal secondary antibody dilution can definitely vary but it might be beneficial to dilute yours further. I noticed that the authors used milk to block, but if the no-protein block is working well then I would recommend continuing to use it.

    I hope this information will be useful, but please let me know if there is anything else that we can do for you, and keep me updated of any progress with the results.

    Read More

    Question

    We bought the 5D2 lipoprotein lipase antibody (catalog # ab93898) but I need to know what is the sequence the antibody binds or at least if it binds at the N or C terminal. Can you provide me with this information?

    Read More
    Answer

    I have found a reference that describes characterization of clone 5D2 (ab93898): Detailed characterization of the binding site of the lipoprotein lipase-specific monoclonal antibody 5D2. Chang et al. J Lipid Research 39:2350-2359, 1998 PMID:9831623 http://www.jlr.org/content/39/12/2350.full.pdf+html In brief, the authors state: "In this study we have shown that the LPL-specific MAb 5D2 recognizes each end of a rather long epitope located between amino acids 380 and 410." This range is closer to the C terminus than the N terminus of the full-length lipase, which consists of 475 amino acids including the signal peptide. http://www.uniprot.org/uniprot/P06858 I hope this is helpful. If you have any other questions, please contact us.

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