Human CPT1A knockout HEK-293T cell lysate (ab256880)
Overview
-
Product name
Human CPT1A knockout HEK-293T cell lysate -
Product overview
Knockout cell lysate achieved by CRISPR/Cas9. -
Parental Cell Line
HEK293T -
Organism
Human -
Mutation description
Knockout achieved by using CRISPR/Cas9, 17 bp deletion in exon 3 and Insertion of the selection cassette in exon 3. -
Passage number
<20 -
Knockout validation
Sanger Sequencing, Western Blot (WB) -
Reconstitution notes
To use as WB control, resuspend the lyophilizate in 50 µL of LDS* Sample Buffer to have a final concentration of 2 mg/ml. For reducing conditions, we recommend a final concentration of 0.1 M DTT.*Usage of SDS sample buffer is not recommended with these lyophilized lysates.
-
Notes
Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version - found here. Please refer to our lysis protocol for further details on how our lysates are prepared.
User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.
Access thousands of knockout cell lysates, generated from commonly used cancer cell lines.
See here for more information on knockout cell lysates.Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.
-
Tested applications
Suitable for: WBmore details
Properties
-
Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab260932 - Human CPT1A knockout HEK293T cell lysate 1 x 100µg ab255553 - Human wild-type HEK293T cell lysate 1 x 100µg -
Research areas
-
Cell type
epithelial -
STR Analysis
Amelogenin X D5S818: 8, 9 D13S317: 12, 14 D7S820: 11 D16S539: 9, 13 vWA: 16, 19 TH01: 7, 9.3 TPOX: 11 CSF1PO: 11, 12
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
-
Alternative names
- Carnitine O palmitoyltransferase 1 liver isoform
- Carnitine O palmitoyltransferase I
- Carnitine O palmitoyltransferase I liver isoform
see all
Associated products
-
KO cell lines
-
Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab256880 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 |
Use at an assay dependent concentration. Predicted molecular weight: 88 kDa.
|
Notes |
---|
WB
Use at an assay dependent concentration. Predicted molecular weight: 88 kDa. |
Images
-
Lane 1: Wild-type HEK-293T cell lysate (20µg)
Lane 2: CPT1A knockout HEK-293T cell lysate (20µg)
Lanes 1- 2: Merged signal (red and green). Green - ab234111 observed at 88 kDa. Red - loading control ab8245 observed at 37 kDa.
ab234111 Anti-CPT1A antibody [EPR21843-71-2F] was shown to specifically 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 and CPT1A knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab234111 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 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. -
Lane 1: Wild-type HEK-293T cell lysate (20µg)
Lane 2: CPT1A knockout HEK-293T cell lysate (20µg)
Lanes 1- 2: Merged signal (red and green). Green - ab220789 observed at 88 kDa. Red - loading control ab8245 observed at 37 kDa.
ab220789 Anti-CPT1A antibody [EPR21843-71-1C] was shown to specifically 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 and CPT1A knockout samples 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) were incubated overnight at 4°C at 1 in 1000 dilution and 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. -
Allele-1: 17 bp deletion in exon 3
-
Allele-2: Insertion of the selection cassette in exon 3
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
-
SDS download
-
Datasheet download
References (0)
ab256880 has not yet been referenced specifically in any publications.