Overview

  • Product name
    Anti-CPT1A antibody [8F6AE9]
    See all CPT1A primary antibodies
  • Description
    Mouse monoclonal [8F6AE9] to CPT1A
  • Host species
    Mouse
  • Tested applications
    Suitable for: Flow Cyt, IHC-P, WB, ICC/IF, In-Cell ELISAmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment, corresponding to amino acids 489-773 of Human CPT1A.

  • Positive control
    • Partial Human Recombinant CPT1A protein; HepG2, H9C2, H4IIE Whole Cell Lysates; Rat heart mitochondrial, Human liver, Rat liver and Mouse liver homogenates; H9C2 cells; HeLa cells IHC-P: human normal kidney FFPE tissue sections
  • General notes

    This antibody clone is manufactured by Abcam.

    This monoclonal antibody to CPT1A has been knockout validated in Western blot. The expected band for CPT1A was observed in wild type cells and the band was not seen in CPT1A knockout cells.

     

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

Properties

Applications

Our Abpromise guarantee covers the use of ab128568 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
Flow Cyt Use at an assay dependent concentration.
IHC-P Use a concentration of 0.5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB Use a concentration of 1 µg/ml. Predicted molecular weight: 88 kDa.
ICC/IF Use a concentration of 0.5 - 5 µg/ml.
In-Cell ELISA Use a concentration of 0.1 - 1 µg/ml.

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

  • Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: CPT1A knockout HAP1 cell lysate (20 µg)
    Lane 3: HeLa cell lysate (20 µg)
    Lane 4: MCF-7 cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab128568 observed at 88 kDa. Red - loading control, ab181602, observed at 37 kDa.


    ab128568 specifically detected the expected band for CPT1A in wild-type HAP1 cells. No band was observed in CPT1A knockout cells. Wild-type and CPT1A knockout samples were subjected to SDS-PAGE. ab128568 and ab181602 (loading control to GAPDH) were diluted at 1 μg/mL and 1/10000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • Immunofluorescent staining of CPT1A in H9C2 cells (rat) using ab128568.
    Fixation: 4% paraformaldehyde PBS fixed for 20 minutes
    Permebilization: 0.1% Triton X-100 PBS for 30 minutes at room temperature while rocking
    Blocking: 2x Sigma Block 0.1% Triton X-100 H2O for 2 hours at room temperature while rocking
    Primary antibodies: Anti-CPT1A antibody (ab128568) 0.5 ug/mL 1x Sigma Block with 0.1% Triton X-100 incubated overnight at 4 °C.
    Washing: 3x 1% NGS 10 minutes/wash.
    Secondary antibodies: Alexa 488 GAM 1:1000 diluted in 1% NGS with 0.1% Triton X-100 PBS incubated for 2 hours at room temperature while rocking.
    Washing: 3x 1% NGS 10 minutes/wash.
    DAPI: 20 ng/mL in 1% NGS, 0.1% Triton X-100 PBS.
    Washing: 1x 1% NGS 10 minutes/wash.
  • All lanes : Anti-CPT1A antibody [8F6AE9] (ab128568) at 1 µg/ml

    Lane 1 : Marker
    Lane 2 : Partial Human Recombinant CPT1A protein (ab128569) at 0.1 µg
    Lane 3 : Full Length Human Recombinant OTC protein at 0.1 µg
    Lane 4 : HepG2 (Human hepatocellular carcinoma cell line) Whole Cell Lysate at 20 µg
    Lane 5 : H9C2 (Rat cardiomyoblast cell line) Whole Cell Lysate at 20 µg
    Lane 6 : H4IIE ( Rat hepatoma cell line) Whole Cell Lysate at 20 µg
    Lane 7 : RHM (Rat heart mitochondrial homogenate) at 20 µg
    Lane 8 : HLH (Human liver homogenate) at 20 µg
    Lane 9 : RLH (Rat liver homogenate) at 20 µg
    Lane 10 : MLH (Mouse liver homogenate) at 20 µg

    Secondary
    All lanes : Goat polyclonal to Mouse IgG – H&L – Pre-Adsorbed (HRP) at 1/10000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

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


    Exposure time: 1 minute


    Predicted partial recombinant protein band size : 32 kDa
    Observed band size : 32 kDa
  • IHC image of CPT1A staining in human normal kidney formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab128568, 0.5µg/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.

    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.
  • Overlay histogram showing HeLa cells stained with ab128568 (red line) or no primary antibody (black line). The cells were fixed with 4% paraformaldehyde (15 min) and then permeabilized with 0.1% Triton X-100 in PBS, 3% BSA for 10 min. The cells were then incubated in 3% BSA in PBS for 10 minutes to block non-specific protein-protein interactions followed by the antibody (ab128568, 1µg/mL) for 60 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96871) at 1/1000 dilution for 30 min.
  • All lanes : Anti-CPT1A antibody [8F6AE9] (ab128568) at 1 µg/ml

    Lane 1 : Marker
    Lane 2 : HHH (Human heart homogenate) RIPA Extract Immunoprecipitated with no primary antibody
    Lane 3 : HHH (Human heart homogenate) RIPA Extract Immunoprecipitated with Anti-CPT1A antibody 8F6AE9 (ab128568)
    Lane 4 : HHH (Human heart homogenate) RIPA Extract 20 µg

    Secondary
    All lanes : Goat polyclonal to Mouse IgG – H&L – Pre-Adsorbed (HRP) at 1:10000

    Developed using the ECL technique.

    Performed under non-reducing conditions.

    Predicted band size: 88 kDa


    Exposure time: 1 minute
  • In-Cell ELISA for Anti-CPT1A antibody (ab128568) stained HeLa cells (human)
    Seeding: HeLa cells seeded in a 1:2 dilution series starting at 60,000 cells/well across Row A, 30,000 cells/well across Row B, etc. Row H contains no cells.
    Table: Example In-Cell ELISA Average Data from shown plate.
    Fixation: 4% paraformaldehyde PBS fixed for 15 minutes
    Permeabilization: 0.3% Triton X-100 PBS for 30 minutes at room temperature while shaking
    Blocking: 2x Sigma Block 0.3% Triton X-100 H2O for 2 hours at room temperature while shaking
    Primary antibodies: All primaries diluted in 1x Sigma Block with 0.3% Triton X-100 incubated overnight at 4 °C.
    • Columns 1-3: Anti-CPT1A antibody (ab128568) 10 ug/mL
    • Columns 4-6: Anti-CPT1A antibody (ab128568) 1 ug/mL
    • Columns 7-9: Anti-CPT1A antibody (ab128568) 0.1 ug/mL
    • Columns 10-12: No Primary
    Washing: Briefly 4x with 0.3% TWEEN-20 PBS

References

This product has been referenced in:
  • Wang C  et al. CPT1A-mediated succinylation of S100A10 increases human gastric cancer invasion. J Cell Mol Med 23:293-305 (2019). Read more (PubMed: 30394687) »
  • Kakimoto PA  et al. Resilient hepatic mitochondrial function and lack of iNOS dependence in diet-induced insulin resistance. PLoS One 14:e0211733 (2019). Read more (PubMed: 30716103) »
See all 43 Publications for this product

Customer reviews and Q&As

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1-4 of 4 Abreviews

Application
Western blot
Sample
Human Tissue lysate - whole (Placenta)
Gel Running Conditions
Reduced Denaturing (10%)
Loading amount
50 µg
Specification
Placenta
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Dr. Polina Vishnyakova

Verified customer

Submitted Apr 02 2019

Application
Western blot
Sample
Human Cell lysate - whole cell (human hepatocytes)
Gel Running Conditions
Reduced Denaturing
Loading amount
15 µg
Specification
human hepatocytes
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Abcam user community

Verified customer

Submitted Jan 25 2018

Application
Western blot
Sample
Horse Cell lysate - other (Liver, mitochondrial extract)
Gel Running Conditions
Reduced Denaturing (12.5 % acrylamide)
Loading amount
60 µg
Specification
Liver, mitochondrial extract
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 7.5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Jul 22 2015

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: RT°C
Sample
Mouse Cell (Mouse alveolar cells)
Specification
Mouse alveolar cells
Permeabilization
Yes - 90% methanol
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Jul 23 2013

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