LDLR KO cell line available to order. KO validated by Next Generation Sequencing. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9; X = 17 bp deletion, 8 bp deletion, 2 bp deletion; Frameshift: 99%.
Images
Key facts
- Cell type
- HeLa
- Species or organism
- Human
- Tissue
- Cervix
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing
- Mutation description
- Knockout achieved by CRISPR/Cas9; X = 17 bp deletion, 8 bp deletion, 2 bp deletion; Frameshift: 99%
Alternative names
F HC, FH, LDL receptor, LDLCQ2, LDLR_HUMAN, Low density lipoprotein receptor class A domain containing protein 3, Low density lipoprotein receptor familial hypercholesterolemia, Low-density lipoprotein receptor
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LDLR KO cell line available to order. KO validated by Next Generation Sequencing. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9; X = 17 bp deletion, 8 bp deletion, 2 bp deletion; Frameshift: 99%.
Key facts
- Cell type
- HeLa
- Species or organism
- Human
- Tissue
- Cervix
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing
- Mutation description
- Knockout achieved by CRISPR/Cas9; X = 17 bp deletion, 8 bp deletion, 2 bp deletion; Frameshift: 99%
- Disease
- Adenocarcinoma
- Concentration
Properties
- Gene name
- LDLR
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Next Generation Sequencing
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 2 US: 2
- Adherent/suspension
- Adherent
- Gender
- Female
Handling procedures
- Initial handling guidelines
Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.
1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.- Subculture guidelines
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 2x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
- Culture medium
- DMEM (High Glucose) + 10% FBS
- Cryopreservation medium
- Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Storage
- Shipped at conditions
- Dry Ice
- Appropriate short-term storage duration
- 1-2 weeks
- Appropriate short-term storage conditions
- -196°C, -80°C
- Appropriate long-term storage conditions
- -196°C
Notes
We will provide viable cells that proliferate on revival.
This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
The LDL Receptor also called LDLR is a protein that plays an important role in the uptake and clearance of low-density lipoproteins (LDL) from the bloodstream. LDLR binds LDL particles facilitating their internalization through receptor-mediated endocytosis. This receptor is expressed mainly in the liver adrenal glands and other tissues involved in cholesterol metabolism. It has an approximate molecular weight of 160 kDa. Researchers often study LDLR using techniques like Western blotting to understand its presence and function.
- Biological function summary
The LDL receptor interacts with LDL particles to regulate cholesterol levels in the body. It is part of a cell surface complex that recognizes and binds to apolipoprotein B-100 or apolipoprotein E present on LDL. This interaction initiates internalization of LDL leading to its degradation in lysosomes where cholesterol can be released and used by the cell. Mutations in the gene encoding LDLR can lead to inefficient cholesterol uptake influencing various metabolic processes.
- Pathways
LDL receptor activities are integral to lipid metabolism and cholesterol homeostasis. Two important biological pathways that involve LDLR include the cholesterol biosynthesis pathway and the lipoprotein clearance pathway. Within these pathways LDLR collaborates closely with proteins like PCSK9 which modulates its expression and degradation and HMG-CoA reductase an important enzyme in cholesterol synthesis to balance cholesterol levels in the body.
- Associated diseases and disorders
Defects in the LDL receptor are strongly associated with familial hypercholesterolemia and atherosclerosis. These conditions arise from impaired clearance of LDL leading to elevated cholesterol levels which pose risks for cardiovascular diseases. LDLR dysfunctions are linked with the Protein PCSK9 whose gain-of-function mutations can exacerbate hypercholesterolemia by promoting degradation of LDLR while statins aim to increase LDLR expression to lower LDL cholesterol levels.
Product promise
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2 product images
Next Generation Sequencing - Human LDLR (LDL Receptor) knockout HeLa cell line (ab273838)
Knockout achieved by CRISPR/Cas9; X = 17 bp deletion, 8 bp deletion, 2 bp deletion; Frameshift: 99%
Western blot - Human LDLR (LDL Receptor) knockout HeLa cell line (ab273838)
Anti-LDLR antibody [EPR24874-56] (Anti-LDL Receptor antibody [EPR24874-56] ab271189) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (Anti-Calnexin antibody [CANX/1543] ab238078) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-LDL Receptor antibody [EPR24874-56] ab271189 was shown to bind specifically to LDLR. A band was observed at 120, 150 kDa in wild-type HeLa cell lysates with no signal observed at this size in LDLR CRISPR-Cas9 edited cell line ab273838 (CRISPR-Cas9 edited cell lysate ab273792). The band observed in the CRISPR-Cas9 edited lysate lane below 120, 150 kDa is likely to represent a possible truncated form of LDLR. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and LDLR CRISPR-Cas9 edited HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween®20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes: Western blot - Anti-LDL Receptor antibody [EPR24874-56] (Anti-LDL Receptor antibody [EPR24874-56] ab271189) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: Western blot - Human LDLR (LDL Receptor) knockout HeLa cell line (ab273838)
Lane 2: LDLR CRISPR-Cas9 edited HeLa cell lysate at 20 µg
Lane 3: HepG2 cell lysate at 20 µg
Lane 4: A431 cell lysate at 20 µg
SecondaryAll lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 95 kDa
Observed band size: 120 kDa, 150 kDa
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