Human SLC27A4 (FATP4) knockout HEK-293T cell line (ab266114)
Overview
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Product name
Human SLC27A4 (FATP4) knockout HEK-293T cell line -
Parental Cell Line
HEK293T -
Organism
Human -
Mutation description
Knockout achieved by using CRISPR/Cas9, Homozygous: 14 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 HEK293T cell line (ab255449). 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.
We will provide viable cells that proliferate on revival.
Properties
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Number of cells
1 x 106 cells/vial, 1 mL -
Adherent /Suspension
Adherent -
Tissue
Kidney -
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 -
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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Relevance
SLC27A4 / FATP4 plays a role in the transport of long chain fatty acids across the plasma membrane. It has acyl-coA ligase activity for long chain and very long chain fatty acids. Deletion of the SLC27A4 gene results in embryonic lethality, which is attributed to the need for fat absorption across the visceral endoderm early in embryonic development. Expression of FAT4P in the intestinal lining is thought to be altered in response to dietary fat. -
Cellular localization
Cell Membrane
Associated products
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KO cell lysates
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab266114 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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WB |
Use at an assay dependent concentration. Predicted molecular weight: 72 kDa.
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Notes |
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WB
Use at an assay dependent concentration. Predicted molecular weight: 72 kDa. |
Images
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All lanes : Anti-SLC27A4 / FATP4 antibody [EPR17319-26] (ab200353) at 1/1000 dilution
Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : SLC27A4 knockout HEK293T cell lysate
Lane 3 : HepG2 cell lysate
Lane 4 : HEK-293 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Predicted band size: 72 kDa
Observed band size: 72 kDaLanes 1-4: Merged signal (red and green). Green - ab200353 observed at 72 kDa. Red - loading control ab8245 observed at 36 kDa.
ab200353 Anti-SLC27A4 / FATP4 antibody [EPR17319-26] was shown to specifically react with SLC27A4 / FATP4 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266114 (knockout cell lysate ab257677) was used. Wild-type and SLC27A4 / FATP4 knockout samples were subjected to SDS-PAGE. ab200353 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.
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All lanes : Anti-SLC27A4 / FATP4 antibody [EPR17319] - C-terminal (ab199719) at 1/1000 dilution
Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : SLC27A4 knockout HEK293T cell lysate
Lane 3 : HepG2 cell lysate
Lane 4 : HEK-293 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Predicted band size: 72 kDa
Observed band size: 72 kDaLanes 1-4: Merged signal (red and green). Green - ab199719 observed at 72 kDa. Red - loading control ab8245 observed at 36 kDa.
ab199719 Anti-SLC27A4 / FATP4 antibody [EPR17319] - C-terminal was shown to specifically react with SLC27A4 / FATP4 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266114 (knockout cell lysate ab257677) was used. Wild-type and SLC27A4 / FATP4 knockout samples were subjected to SDS-PAGE. ab199719 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.
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Homozygous: 14 bp deletion in exon 2
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Representative images of SLC27A4 knockout HEK293T cells, low and high confluency examples (top left and right respectively) and wild-type HEK293T cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using an EVOS M5000 microscope.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (0)
ab266114 has not yet been referenced specifically in any publications.