Recombinant
RabMAb

Anti-Adipose Triglyceride Lipase antibody [EPR19650] (ab207799)

Overview

  • Product name
    Anti-Adipose Triglyceride Lipase antibody [EPR19650]
    See all Adipose Triglyceride Lipase primary antibodies
  • Description
    Rabbit monoclonal [EPR19650] to Adipose Triglyceride Lipase
  • Tested applications
    Suitable for: WB, IHC-P, ICC/IF, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Human Adipose Triglyceride Lipase aa 300 to the C-terminus. The exact sequence is proprietary.
    Database link: Q96AD5

  • Positive control
    • WB: Human adipose tissue lysate; Adult mouse and rat adipose tissue lysates. IHC-P: Human Adipose tissue; Mouse and rat white and brown adipose tissue. ICC/IF: 3T3-L1 cells. IP: 3T3-L1 differentiated for 6 days whole cell lysate.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab207799 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 1/1000. Detects a band of approximately 55 kDa (predicted molecular weight: 55 kDa).

Abcam recommends milk blocking for this product.

IHC-P 1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/500.
IP 1/30.

Target

  • Function
    Catalyzes the initial step in triglyceride hydrolysis in adipocyte and non-adipocyte lipid droplets. Also has acylglycerol transacylase activity. May act coordinately with LIPE/HLS within the lipolytic cascade. Regulates adiposome size and may be involved in the degradation of adiposomes. May play an important role in energy homeostasis. May play a role in the response of the organism to starvation, enhancing hydrolysis of triglycerides and providing free fatty acids to other tissues to be oxidized in situations of energy depletion.
  • Tissue specificity
    Highest expression in adipose tissue. Also detected in heart, skeletal muscle, and portions of the gastrointestinal tract. Detected in normal retina and retinoblastoma cells. Detected in retinal pigment epithelium and, at lower intensity, in the inner segments of photoreceptors and in the ganglion cell layer of the neural retina (at protein level).
  • Pathway
    Glycerolipid metabolism; triacylglycerol degradation.
  • Involvement in disease
    Note=Genetic variations in PNPLA2 may be associated with risk of diabetes mellitus type 2.
    Defects in PNPLA2 are the cause of neutral lipid storage disease with myopathy (NLSDM) [MIM:610717]; also known as neutral lipid storage disease without ichthyosis. NSLDM is a neutral lipid storage disorder (NLSD) with myopathy but without ichthyosis. NLSDs are characterized by the presence of triglyceride-containing cytoplasmic droplets in leukocytes and in other tissues, including bone marrow, skin, and muscle. Individuals with NLSDM did not show obesity, in spite of a defect in triglyceride degradation in fibroblasts and in marked triglyceride storage in liver, muscles, and other visceral cells.
  • Sequence similarities
    Contains 1 patatin domain.
  • Developmental stage
    Induced during differentiation of primary preadipocytes to adipocytes. Expression increased from fetal to adult in retinal pigment epithelium.
  • Cellular localization
    Lipid droplet. Cell membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • 1110001C14Rik antibody
    • Adipose triglyceride lipase antibody
    • ATGL antibody
    • ATGL DESNUTRIN antibody
    • Calcium independent phospholipase A2 antibody
    • Calcium-independent phospholipase A2 antibody
    • Desnutrin antibody
    • EC 3.1.1.3 antibody
    • FP17548 antibody
    • IPLA2 zeta antibody
    • IPLA2-zeta antibody
    • Mutant patatin like phospholipase domain containing 2 antibody
    • Patatin like phospholipase domain containing 2 antibody
    • PATATIN LIKE PHOSPHOLIPASE DOMAIN CONTAINING PROTEIN 2 antibody
    • Patatin-like phospholipase domain-containing protein 2 antibody
    • PEDF R antibody
    • PHOSPHOLIPASE A2 CALCIUM INDEPENDENT ZETA antibody
    • Pigment epithelium derived factor antibody
    • Pigment epithelium-derived factor antibody
    • plpl antibody
    • plpl2 antibody
    • PLPL2_HUMAN antibody
    • Pnpla2 antibody
    • Transport secretion protein 2 antibody
    • Transport secretion protein 2.2 antibody
    • Transport-secretion protein 2 antibody
    • Triglyceride hydrolase antibody
    • TTS 2.2 antibody
    • TTS2 antibody
    • TTS2.2 antibody
    • ZETA antibody
    see all

Images

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized 3T3-L1 (mouse embryonic fibroblast cell line) undifferentiated and differentiated cells labeling Adipose Triglyceride Lipase with ab207799 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing positive staining on 3T3-L1 cells differentiated for 6 days. The level of expression in 3T3/L1 can be induced by differentiation treatment according to the literature (PMID 19297333).

    The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

  • Immunohistochemical analysis of paraffin-embedded rat brown adipose tissue labeling Adipose Triglyceride Lipase with ab207799 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on rat brown adipose tissue is observed (PMID: 15550674). Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ab97051 at 1/500 dilution.

  • Immunohistochemical analysis of paraffin-embedded rat white adipose tissue labeling Adipose Triglyceride Lipase with ab207799 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on rat white adipose tissue is observed (PMID: 15550674). Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ab97051 at 1/500 dilution.

  • Immunohistochemical analysis of paraffin-embedded mouse brown adipose tissue labeling Adipose Triglyceride Lipase with ab207799 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on mouse brown adipose tissue is observed (PMID: 15550674). Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ab97051 at 1/500 dilution.

  • Immunohistochemical analysis of paraffin-embedded mouse white adipose tissue labeling Adipose Triglyceride Lipase with ab207799 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on mouse white adipose tissue is observed (PMID: 15550674). Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ab97051 at 1/500 dilution.

  • Adipose Triglyceride Lipase was immunoprecipitated from 0.35 mg of 3T3-L1 (mouse embryonic fibroblast cell line) differentiated for 6 days whole cell lysate with ab207799 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab207799 at 1/500 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366), was used as secondary antibody at 1/1000 dilution.


    Lane 1: 3T3-L1 differentiated for 6 days whole cell lysate 10 µg (Input).

    Lane 2: ab207799 IP in 3T3-L1 differentiated for 6 days whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab207799 in 3T3-L1 differentiated for 6 days whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.
    Exposure time: 1 second.

  • IHC image of Adipose Triglyceride Lipase staining in a formalin-fixed, paraffin-embedded human adipose tissue section, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval (EDTA based pH 9.0 solution, epitope retrieval solution 2) for 20 mins. The section was then incubated with ab207799, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. As a negative control (inset), an identical assay was performed without adding the primary antibody.

     

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • Lane 1 : Anti-Adipose Triglyceride Lipase antibody [EPR19650] (ab207799) at 1/1000 dilution (2% Bovine Serum Albumin)
    Lane 2 : Anti-Adipose Triglyceride Lipase antibody [EPR19650] (ab207799) at 1/1000 dilution (3% Milk)

    Lane 1 : Human adipose normal tissue lysate - total protein (ab28980)
    Lane 2 : Human adipose normal tissue lysate - total protein (ab28980)

    Lysates/proteins at 10 µg per lane.

    Secondary
    Peroxidase AffiniPure Goat Anti-Rabbit IgG (H+L) at 1/50000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 55 kDa
    Observed band size : 55 kDa


    Exposure time : 8 minutes

    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin (lane 1) and 3% Milk (lane 2) before being incubated with ab207799 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.

  • All lanes : Anti-Adipose Triglyceride Lipase antibody [EPR19650] (ab207799) at 1/1000 dilution (3% Milk)

    Lane 1 : Adult Mouse Adipose Tissue Lysate
    Lane 2 : Adult Rat Adipose Tissue Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Peroxidase AffiniPure Goat Anti-Rabbit IgG (H+L) at 1/50000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 55 kDa
    Observed band size : 55 kDa


    Exposure time : 5 seconds

    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% Milk before being incubated with ab207799 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.

References

ab207799 has not yet been referenced specifically in any publications.

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