Anti-CPT1A antibody [8F6AE9] (ab128568)
Key features and details
- Mouse monoclonal [8F6AE9] to CPT1A
- Suitable for: Flow Cyt, IHC-P, WB, ICC/IF, In-Cell ELISA
- Knockout validated
- Reacts with: Mouse, Rat, Human
- Isotype: IgG2b
Related conjugates and formulations
Overview
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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.
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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
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General notes
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.
This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.
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 this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. 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, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.4
Preservative: 0.02% Sodium azide
Constituent: HEPES buffered saline -
Concentration information loading...
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Purity
Ammonium Sulphate Precipitation -
Purification notes
Purity is near homogeneity as judged by SDS-PAGE. The antibody was produced in vitro using hybridomas grown in serum-free medium, and then concentrated by ammonium sulfate precipitation. -
Clonality
Monoclonal -
Clone number
8F6AE9 -
Isotype
IgG2b -
Light chain type
kappa -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Positive Controls
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Related Products
Applications
The Abpromise guarantee
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 |
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Flow Cyt |
Use at an assay dependent concentration.
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IHC-P | (1) |
Use a concentration of 0.5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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WB | (3) |
Use a concentration of 1 µg/ml. Predicted molecular weight: 88 kDa.
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ICC/IF | (1) |
Use a concentration of 0.5 - 5 µg/ml.
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In-Cell ELISA |
Use a concentration of 0.1 - 1 µg/ml.
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Notes |
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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
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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
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Database links
- Entrez Gene: 1374 Human
- Entrez Gene: 12894 Mouse
- Entrez Gene: 25757 Rat
- Omim: 600528 Human
- SwissProt: P50416 Human
- SwissProt: P97742 Mouse
- SwissProt: P32198 Rat
- Unigene: 503043 Human
see all -
Alternative names
- Carnitine O palmitoyltransferase 1 liver isoform antibody
- Carnitine O palmitoyltransferase I antibody
- Carnitine O palmitoyltransferase I liver isoform antibody
see all
Images
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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 -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CPT1A antibody [8F6AE9] (ab128568)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.
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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
Datasheets and documents
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SDS download
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Datasheet download
References (115)
ab128568 has been referenced in 115 publications.
- Wang L et al. Engineered Bacteria of MG1363-pMG36e-GLP-1 Attenuated Obesity-Induced by High Fat Diet in Mice. Front Cell Infect Microbiol 11:595575 (2021). PubMed: 33732656
- Luo P et al. S-Allylmercaptocysteine improves alcoholic liver disease partly through a direct modulation of insulin receptor signaling. Acta Pharm Sin B 11:668-679 (2021). PubMed: 33777674
- Mori H et al. The molecular and metabolic program by which white adipocytes adapt to cool physiologic temperatures. PLoS Biol 19:e3000988 (2021). PubMed: 33979328
- Miguel V et al. The program of renal fibrogenesis is controlled by microRNAs regulating oxidative metabolism. Redox Biol 40:101851 (2021). PubMed: 33465566
- Tokoro M et al. a-Tocopherol suppresses hepatic steatosis by increasing CPT-1 expression in a mouse model of diet-induced nonalcoholic fatty liver disease. Obes Sci Pract 7:91-99 (2021). PubMed: 33680496