Human SCARB1 knockout HEK-293T cell line (ab282646)
Overview
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Product name
Human SCARB1 knockout HEK-293T cell line
See all Scavenging Receptor SR-BI lysates -
Parental Cell Line
HEK293T -
Organism
Human -
Mutation description
Knockout achieved by using CRISPR/Cas9, Homozygous: 40 bp deletion in exon 2 -
Passage number
<20 -
Knockout validation
Sanger Sequencing, Western Blot (WB) -
Tested applications
Suitable for: WBmore details -
Biosafety level
2 -
General notes
Recommended control: Human wild-type HEK-293T cell line (ab282205). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.
Cryopreservation cell medium: Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Culture medium: DMEM (High Glucose) + 10% FBS
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. Based on cell count, seed cells in an appropriate cell culture 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.
4. Incubate the culture at 37°C incubator with 5% CO2. 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.
- A partial media change 24 hours prior to subculture may be helpful to encourage growth, if required.
- Cells should be passaged when they have achieved 80-90% confluence.
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.
Properties
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Number of cells
1 x 106 cells/vial, 1 mL -
Viability
~80% -
Adherent /Suspension
Adherent -
Tissue
Kidney -
Cell type
epithelial -
Gender
Female -
Mycoplasma free
Yes -
Storage instructions
Shipped on Dry Ice. Store in liquid nitrogen. -
Storage buffer
Constituents: 8.7% Dimethylsulfoxide, 2% Cellulose, methyl ether -
Research areas
Target
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Function
Receptor for different ligands such as phospholipids, cholesterol ester, lipoproteins, phosphatidylserine and apoptotic cells. Probable receptor for HDL, located in particular region of the plasma membrane, called caveolae. Facilitates the flux of free and esterified cholesterol between the cell surface and extracellular donors and acceptors, such as HDL and to a lesser extent, apoB-containing lipoproteins and modified lipoproteins. Probably involved in the phagocytosis of apoptotic cells, via its phosphatidylserine binding activity. Receptor for hepatitis C virus glycoprotein E2. Binding between SCARB1 and E2 was found to be independent of the genotype of the viral isolate. Plays an important role in the uptake of HDL cholesteryl ester. -
Tissue specificity
Widely expressed. -
Sequence similarities
Belongs to the CD36 family. -
Post-translational
modificationsN-glycosylated. -
Cellular localization
Cell membrane. Membrane > caveola. Predominantly localized to cholesterol and sphingomyelin-enriched domains within the plasma membrane, called caveolae. - Information by UniProt
Associated products
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KO cell lysates
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab282646 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.
|
| Notes |
|---|
|
WB
Use at an assay dependent concentration. |
Images
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Western blot - Human SCARB1 knockout HEK-293T cell line (ab282646)All lanes : Anti-Scavenging Receptor SR-BI antibody [25/CLA-1] (ab300632) at 1/1000 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : SCARB1 knockout HEK-293T cell lysate
Lane 3 : Human Liver cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Observed band size: 70,75 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-Scavenging Receptor SR-BI antibody [25/CLA-1] staining at 1/1000 dilution, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab300632 was shown to bind specifically to Scavenging Receptor SR-BI. A band was observed at 70 and 75 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in SCARB1 knockout cell line ab282646. To generate this image, wild-type and SCARB1 knockout HEK-293T cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % 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-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) at 1/20000 dilution.
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Western blot - Human SCARB1 knockout HEK-293T cell line (ab282646)All lanes : Anti-Scavenging Receptor SR-BI antibody [EPR20190] (ab217318) at 1/2000 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : SCARB1 knockout HEK-293T cell lysate
Lane 3 : Human Liver cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Observed band size: 70,75 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-Scavenging Receptor SR-BI antibody [EPR20190] staining at 1/2000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab217318 was shown to bind specifically to Scavenging Receptor SR-BI. A band was observed at 70/75 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in SCARB1 knockout cell line ab282646 (knockout cell lysate ab283046). To generate this image, wild-type and SCARB1 knockout HEK-293T cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % 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 (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
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Western blot - Human SCARB1 knockout HEK-293T cell line (ab282646)All lanes : Anti-Scavenging Receptor SR-BI antibody [EP1556Y] (ab52629) at 1/1000 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : SCARB1 knockout HEK-293T cell lysate
Lane 3 : Human Liver cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Observed band size: 70,75 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-Scavenging Receptor SR-BI antibody [EP1556Y] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab52629 was shown to bind specifically to Scavenging Receptor SR-BI. A band was observed at 70/75 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in SCARB1 knockout cell line ab282646 (knockout cell lysate ab283046). To generate this image, wild-type and SCARB1 knockout HEK-293T cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % 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 (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
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Sanger Sequencing - Human SCARB1 knockout HEK-293T cell line (ab282646)40 bp deletion in exon 2
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
References (0)
ab282646 has not yet been referenced specifically in any publications.