Human LTA4H (Leukotriene A4 hydrolase) knockout HEK-293T cell line (ab266467)
Overview
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Product name
Human LTA4H (Leukotriene A4 hydrolase) knockout HEK-293T cell line -
Parental Cell Line
HEK293T -
Organism
Human -
Mutation description
Knockout achieved by using CRISPR/Cas9, Homozygous: 10 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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Function
Hydrolyzes an epoxide moiety of leukotriene A4 (LTA-4) to form leukotriene B4 (LTB-4). The enzyme also has some peptidase activity. -
Pathway
Lipid metabolism; leukotriene B4 biosynthesis. -
Sequence similarities
Belongs to the peptidase M1 family. -
Cellular localization
Cytoplasm. - Information by UniProt
Associated products
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KO cell lysates
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Related Products
- Anti-Leukotriene A4 hydrolase/LTA4H antibody [EPR5712] (ab109434)
- Anti-Leukotriene A4 hydrolase/LTA4H antibody [EPR5713] (ab133512)
- Anti-Leukotriene A4 hydrolase/LTA4H antibody [EPR5713] - BSA and Azide free (ab240064)
- Anti-Leukotriene A4 hydrolase/LTA4H antibody [EPR5712] - BSA and Azide free (ab247861)
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab266467 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: 69 kDa.
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Notes |
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WB
Use at an assay dependent concentration. Predicted molecular weight: 69 kDa. |
Images
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All lanes : Anti-Leukotriene A4 hydrolase/LTA4H antibody [EPR5712] (ab109434) at 1/1000 dilution
Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : LTA4H knockout HEK293T cell lysate
Lane 3 : T-47D cell lysate
Lane 4 : A549 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: 69 kDa
Observed band size: 69 kDaLanes 1-4: Merged signal (red and green). Green - ab109434 observed at 69 kDa. Red - loading control ab8245 observed at 36 kDa.
ab109434 Anti-Leukotriene A4 hydrolase/LTA4H antibody [EPR5712] was shown to specifically react with Leukotriene A4 hydrolase/LTA4H in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266467 (knockout cell lysate ab258034) was used. Wild-type and Leukotriene A4 hydrolase/LTA4H knockout samples were subjected to SDS-PAGE. ab109434 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-Leukotriene A4 hydrolase/LTA4H antibody [EPR5713] (ab133512) at 1/1000 dilution
Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : LTA4H knockout HEK293T cell lysate
Lane 3 : T-47D cell lysate
Lane 4 : A549 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: 69 kDa
Observed band size: 69 kDaLanes 1-4: Merged signal (red and green). Green - ab133512 observed at 69 kDa. Red - loading control ab8245 observed at 36 kDa.
ab133512 Anti-Leukotriene A4 hydrolase/LTA4H antibody [EPR5713] was shown to specifically react with Leukotriene A4 hydrolase/LTA4H in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266467 (knockout cell lysate ab258034) was used. Wild-type and Leukotriene A4 hydrolase/LTA4H knockout samples were subjected to SDS-PAGE. ab133512 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: 10 bp deletion in exon 2
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (0)
ab266467 has not yet been referenced specifically in any publications.