Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-CPT1A antibody [EPR21843-71-1C] (ab220789)

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Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR21843-71-1C] to CPT1A
  • Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IP
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-CPT1A antibody [EPR21843-71-1C]
    See all CPT1A primary antibodies

  • Description

    Rabbit monoclonal [EPR21843-71-1C] to CPT1A

  • Host species

    Rabbit

  • Tested applications

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

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Recombinant fragment within Human CPT1A aa 400 to the C-terminus. The exact sequence is proprietary.
    Database link: P50416

  • Positive control

    • WB: Wild-type HAP1 whole cell lysate; HEK-293T, HeLa, SK-OV-3, MCF7 and HepG2 whole cell lysates; Human kidney lysate; His-tagged human CPT1A recombinant protein (aa406-755). IHC-P: Human kidney and ovarian carcinoma tissues. ICC/IF: HeLa and SK-OV-3 cells. Flow Cyt: HeLa cells. IP: SK-OV-3 whole cell lysate.

  • 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.

    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 ab220789 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 88 kDa (predicted molecular weight: 88 kDa).
IHC-P 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/100.
Flow Cyt 1/600.
IP 1/30.

Target

  • Tissue specificity

    Strong expression in kidney and heart, and lower in liver and skeletal muscle.

  • Pathway

    Lipid metabolism; fatty acid beta-oxidation.

  • Involvement in disease

    Defects in CPT1A are the cause of carnitine palmitoyltransferase 1A deficiency (CPT1AD) [MIM:255120]; also known as CPT-I deficiency or CPT1A deficiency. CPT1AD is a rare autosomal recessive metabolic disorder of long-chain fatty acid oxidation characterized by severe episodes of hypoketotic hypoglycemia usually occurring after fasting or illness. Onset is in infancy or early childhood.

  • Sequence similarities

    Belongs to the carnitine/choline acetyltransferase family.

  • Cellular localization

    Mitochondrion outer membrane.
  • Information by UniProt

  • Database links

    • Alternative names

      • Carnitine O palmitoyltransferase 1 liver isoform antibody
      • Carnitine O palmitoyltransferase I antibody
      • Carnitine O palmitoyltransferase I liver isoform antibody
      • Carnitine O-palmitoyltransferase 1 antibody
      • Carnitine O-palmitoyltransferase I antibody
      • Carnitine palmitoyltransferase 1A (liver) antibody
      • Carnitine palmitoyltransferase 1A antibody
      • Carnitine palmitoyltransferase I antibody
      • Carnitine palmitoyltransferase I liver antibody
      • CPT 1 antibody
      • CPT I antibody
      • CPT1 antibody
      • CPT1 L antibody
      • CPT1-L antibody
      • Cpt1a antibody
      • CPT1A_HUMAN antibody
      • CPTI antibody
      • CPTI-L antibody
      • L CPT1 antibody
      • liver isoform antibody
      see all

    Images

    • Western blot - Anti-CPT1A antibody [EPR21843-71-1C] (ab220789)
      Western blot - Anti-CPT1A antibody [EPR21843-71-1C] (ab220789)
      All lanes : Anti-CPT1A antibody [EPR21843-71-1C] (ab220789) at 1/1000 dilution

      Lane 1 : Wild-type HEK-293T cell lysate
      Lane 2 : CPT1A knockout HEK-293T cell lysate
      Lane 3 : Wild-type HAP1 cell lysate
      Lane 4 : CPT1A knockout HAP1 cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 88 kDa



      Lanes 1-4: Merged signal (red and green). Green - ab220789 observed at 88 kDa. Red - loading control ab8245 observed at 37 kDa.

       ab220789 Anti-CPT1A antibody [EPR21843-71-1C] was shown to specifically react with CPT1A in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266319 (knockout cell lysate ab256880) was used. Wild-type and CPT1A knockout samples were subjected to SDS-PAGE. ab220789 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.

       

    • Western blot - Anti-CPT1A antibody [EPR21843-71-1C] (ab220789)
      Western blot - Anti-CPT1A antibody [EPR21843-71-1C] (ab220789)
      All lanes : Anti-CPT1A antibody [EPR21843-71-1C] (ab220789) at 1/5000 dilution

      Lane 1 : Wild-type HAP1 whole cell lysate
      Lane 2 : CPT1A knockout HAP1 whole cell lysate
      Lane 3 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
      Lane 4 : SK-OV-3 (human ovarian cancer cell line) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Developed using the ECL technique.

      Predicted band size: 88 kDa
      Observed band size: 88 kDa


      Exposure time: 92 seconds


      Blocking/Dilution buffer: 5% NFDM/TBST.

      ab220789 was shown to specifically react with CPT1A in wild-type HAP1 cells as signal was lost in CPT1A knockout cells. Wild-type and CPT1A knockout samples were subjected to SDS-PAGE. ab220789 and ab181602 (Rabbit anti-GAPDH loading control) were incubated 1 hour at room temperature at 1/5000 dilution and 1/200,000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/100,000 dilution for 1 hour at room temperature before imaging. The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument using the ECL technique.

    • Immunocytochemistry/ Immunofluorescence - Anti-CPT1A antibody [EPR21843-71-1C] (ab220789)
      Immunocytochemistry/ Immunofluorescence - Anti-CPT1A antibody [EPR21843-71-1C] (ab220789)

      Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized SK-OV-3 (human ovarian cancer cell line) cells labeling CPT1A with ab220789 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing mitochondrial staining in SK-OV-3 cell line.

      The nuclear counter stain is DAPI (blue). COX IV is detected with Anti-COX IV antibody [mAbcam33985] - Mitochondrial Marker (ab33985) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) (red).

      The negative controls are as follows:

      -ve control 1: ab220789 at 1/100 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) secondary antibody at 1/1000 dilution.

      -ve control 2: Anti-COX IV antibody [mAbcam33985] - Mitochondrial Marker (ab33985) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CPT1A antibody [EPR21843-71-1C] (ab220789)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CPT1A antibody [EPR21843-71-1C] (ab220789)

      Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling CPT1A with ab220789 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Granular cytoplasmic staining in human kidney (PMID: 18192268; PMID: 28956034). Counter stained with hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunoprecipitation - Anti-CPT1A antibody [EPR21843-71-1C] (ab220789)
      Immunoprecipitation - Anti-CPT1A antibody [EPR21843-71-1C] (ab220789)

      CPT1A was immunoprecipitated from 0.35 mg of SK-OV-3 (human ovarian cancer cell line) whole cell lysate with ab220789 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab220789 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

      Lane 1: SK-OV-3 whole cell lysate 10 µg (Input). 

      Lane 2: ab220789 IP in SK-OV-3 whole cell lysate. 

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab220789 in SK-OV-3 whole cell lysate.

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

      Exposure time: 30 seconds.

    • Flow Cytometry - Anti-CPT1A antibody [EPR21843-71-1C] (ab220789)
      Flow Cytometry - Anti-CPT1A antibody [EPR21843-71-1C] (ab220789)

      Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cell line labeling CPT1A with ab220789 at 1/600 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CPT1A antibody [EPR21843-71-1C] (ab220789)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CPT1A antibody [EPR21843-71-1C] (ab220789)

      Immunohistochemical analysis of paraffin-embedded human ovarian carcinoma tissue labeling CPT1A with ab220789 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Granular cytoplasmic staining in human ovarian carcinoma (PMID: 26716645). Counter stained with hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunocytochemistry/ Immunofluorescence - Anti-CPT1A antibody [EPR21843-71-1C] (ab220789)
      Immunocytochemistry/ Immunofluorescence - Anti-CPT1A antibody [EPR21843-71-1C] (ab220789)

      Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling CPT1A with ab220789 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing mitochondrial staining in HeLa cell line.

      The nuclear counter stain is DAPI (blue). COX IV is detected with Anti-COX IV antibody [mAbcam33985] - Mitochondrial Marker (ab33985) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) (red).

      The negative controls are as follows:

      -ve control 1: ab220789 at 1/100 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) secondary antibody at 1/1000 dilution.

      -ve control 2: Anti-COX IV antibody [mAbcam33985] - Mitochondrial Marker (ab33985) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077).

    • Western blot - Anti-CPT1A antibody [EPR21843-71-1C] (ab220789)
      Western blot - Anti-CPT1A antibody [EPR21843-71-1C] (ab220789)
      All lanes : Anti-CPT1A antibody [EPR21843-71-1C] (ab220789) at 1/1000 dilution

      Lane 1 : MCF7 (human breast adenocarcinoma cell line) whole cell lysate
      Lane 2 : HepG2 (human liver hepatocellular carcinoma cell line) whole cell lysate
      Lane 3 : Human kidney lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution

      Developed using the ECL technique.

      Predicted band size: 88 kDa
      Observed band size: 88 kDa



      Exposure time : Lane 1: 3 minutes; Lane 2: 8 seconds; Lane 3: 10 seconds.

      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Western blot - Anti-CPT1A antibody [EPR21843-71-1C] (ab220789)
      Western blot - Anti-CPT1A antibody [EPR21843-71-1C] (ab220789)
      All lanes : Anti-CPT1A antibody [EPR21843-71-1C] (ab220789) at 1/1000 dilution

      Lane 1 : His-tagged human CPT1A recombinant protein (aa406-755)
      Lane 2 : His-tagged human CPT1B recombinant protein (aa407-756)

      Lysates/proteins at 0.02 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Developed using the ECL technique.

      Predicted band size: 88 kDa
      Observed band size: 38 kDa why is the actual band size different from the predicted?


      Exposure time: 10 seconds


      Blocking/Dilution buffer: 5% NFDM/TBST.

    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.

    Click here to view the general protocols

    References (0)

    Publishing research using ab220789? Please let us know so that we can cite the reference in this datasheet.

    ab220789 has not yet been referenced specifically in any publications.

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