Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-CPT1A antibody [EPR21843-71-1C] - BSA and Azide free (ab234906)

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 - BSA and Azide free
  • Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IP
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-CPT1A antibody [EPR21843-71-1C] - BSA and Azide free
    See all CPT1A primary antibodies

  • Description

    Rabbit monoclonal [EPR21843-71-1C] to CPT1A - BSA and Azide free

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    ICC/IF
    Human
    IHC-P
    Human
    IP
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • 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

    Ab234906 is the carrier-free version of ab220789. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with

    ab234906 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Images

  • Western blot - Anti-CPT1A antibody [EPR21843-71-1C] - BSA and Azide free (ab234906)
    Western blot - Anti-CPT1A antibody [EPR21843-71-1C] - BSA and Azide free (ab234906)
    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

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

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



    This data was developed using the same antibody clone in a different buffer formulation (ab220789).

      Lanes 1- 2: Merged signal (red and green). Green - ab220789 observed at 88 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

     ab220789 was shown to react with CPT1A in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266319 (knockout cell lysate ab256880) was used. Wild-type HEK-293T and CPT1A knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab220789 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CPT1A antibody [EPR21843-71-1C] - BSA and Azide free (ab234906)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CPT1A antibody [EPR21843-71-1C] - BSA and Azide free (ab234906)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab220789).

    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] - BSA and Azide free (ab234906)
    Immunocytochemistry/ Immunofluorescence - Anti-CPT1A antibody [EPR21843-71-1C] - BSA and Azide free (ab234906)

    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).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab220789).

  • Western blot - Anti-CPT1A antibody [EPR21843-71-1C] - BSA and Azide free (ab234906)
    Western blot - Anti-CPT1A antibody [EPR21843-71-1C] - BSA and Azide free (ab234906)
    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

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab220789).

  • Immunoprecipitation - Anti-CPT1A antibody [EPR21843-71-1C] - BSA and Azide free (ab234906)
    Immunoprecipitation - Anti-CPT1A antibody [EPR21843-71-1C] - BSA and Azide free (ab234906)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab220789).

  • Flow Cytometry - Anti-CPT1A antibody [EPR21843-71-1C] - BSA and Azide free (ab234906)
    Flow Cytometry - Anti-CPT1A antibody [EPR21843-71-1C] - BSA and Azide free (ab234906)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab220789).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CPT1A antibody [EPR21843-71-1C] - BSA and Azide free (ab234906)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CPT1A antibody [EPR21843-71-1C] - BSA and Azide free (ab234906)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab220789).

    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] - BSA and Azide free (ab234906)
    Immunocytochemistry/ Immunofluorescence - Anti-CPT1A antibody [EPR21843-71-1C] - BSA and Azide free (ab234906)

    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).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab220789).

  • Anti-CPT1A antibody [EPR21843-71-1C] - BSA and Azide free (ab234906)
    Anti-CPT1A antibody [EPR21843-71-1C] - BSA and Azide free (ab234906)

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