Validated using a knockout cell line
Recombinant
RabMAb

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

Knockout Tested Rabbit recombinant monoclonal CPT1A antibody [EPR21843-71-1C]. Validated in WB, IP, IHC, Flow Cyt, ICC/IF and tested in Human. Immunogen corresponding to recombinant fragment.

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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; 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

    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.

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 1/5000 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.

      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.

      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.

    References

    ab220789 has not yet been referenced specifically in any publications.

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

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