Recombinant Anti-CPT1A antibody [EPR21843-71-1C] (ab220789)
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: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IP
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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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: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IPmore details -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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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 (intra): HeLa cells. IP: SK-OV-3 whole cell lysate.
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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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR21843-71-1C -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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KO cell pellets
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Related Products
Applications
The Abpromise guarantee
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 |
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Flow Cyt (Intra) |
1/600.
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WB |
1/1000. Detects a band of approximately 88 kDa (predicted molecular weight: 88 kDa).
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IHC-P |
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF |
1/100.
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IP |
1/30.
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Notes |
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Flow Cyt (Intra)
1/600. |
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. |
IP
1/30. |
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
- Omim: 600528 Human
- SwissProt: P50416 Human
- Unigene: 503043 Human
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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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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 kDaLanes 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.
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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).
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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.
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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.
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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 secondsBlocking/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.
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Intracellular 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.
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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.
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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).
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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 kDaExposure time : Lane 1: 3 minutes; Lane 2: 8 seconds; Lane 3: 10 seconds.
Blocking/Dilution buffer: 5% NFDM/TBST.
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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 secondsBlocking/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.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (1)
ab220789 has been referenced in 1 publication.
- Sun C et al. Mass spectrometry imaging-based metabolomics to visualize the spatially resolved reprogramming of carnitine metabolism in breast cancer. Theranostics 10:7070-7082 (2020). PubMed: 32641979