Human LTA4H (Leukotriene A4 hydrolase) knockout HEK-293T cell line
- Advanced Validation
- What is this?
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LTA4H KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 10 bp deletion in exon 2.
View Alternative Names
FLJ17564, LKHA4_HUMAN, LTA-4 hydrolase, LTA4, LTA4H, Leukotriene A(4) hydrolase, Leukotriene A-4 hydrolase
- WB
Lab
Western blot - Human LTA4H (Leukotriene A4 hydrolase) knockout HEK-293T cell line (AB266467)
Lanes 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.
All lanes:
Western blot - Anti-Leukotriene A4 hydrolase/LTA4H antibody [EPR5712] (<a href='/en-us/products/primary-antibodies/leukotriene-a4-hydrolase-lta4h-antibody-epr5712-ab109434'>ab109434</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK293T cell lysate at 20 µg
Lane 2:
LTA4H knockout HEK293T cell lysate at 20 µg
Lane 2:
Western blot - Human LTA4H (Leukotriene A4 hydrolase) knockout HEK-293T cell line (ab266467)
Lane 3:
T-47D cell lysate at 20 µg
Lane 4:
A549 cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution
Predicted band size: 19 kDa,40 kDa,44 kDa,47 kDa,69 kDa,80 kDa
Observed band size: 100 kDa,20 kDa,40 kDa,50 kDa,60 kDa,69 kDa
false
- WB
Lab
Western blot - Human LTA4H (Leukotriene A4 hydrolase) knockout HEK-293T cell line (AB266467)
Lanes 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.
All lanes:
Western blot - Anti-Leukotriene A4 hydrolase/LTA4H antibody [EPR5713] (<a href='/en-us/products/primary-antibodies/leukotriene-a4-hydrolase-lta4h-antibody-epr5713-ab133512'>ab133512</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK293T cell lysate at 20 µg
Lane 2:
LTA4H knockout HEK293T cell lysate at 20 µg
Lane 2:
Western blot - Human LTA4H (Leukotriene A4 hydrolase) knockout HEK-293T cell line (ab266467)
Lane 3:
T-47D cell lysate at 20 µg
Lane 4:
A549 cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution
Predicted band size: 69 kDa
Observed band size: 69 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human LTA4H (Leukotriene A4 hydrolase) knockout HEK-293T cell line (AB266467)
Homozygous : 10 bp deletion in exon 2
- Cell Culture
Lab
Cell Culture - Human LTA4H (Leukotriene A4 hydrolase) knockout HEK-293T cell line (AB266467)
Representative images LTA4H 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 a EVOS M5000 microscope.
Reactivity data
Product details
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.
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.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Zygosity
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
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.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
Leukotriene A4 hydrolase (LTA4H) transforms leukotriene A4 an unstable epoxide into leukotriene B4 a potent chemoattractant. The enzyme does not function within a multi-protein complex but interacts with small molecules during leukotriene biosynthesis. Leukotriene B4 produced by LTA4H recruits neutrophils to sites of inflammation enhancing immune defense. While its primary function is promoting inflammation this activity also regulates the resolution phase by facilitating tissue repair and return to homeostasis.
Pathways
Leukotriene A4 hydrolase plays a role in the arachidonic acid cascade an important biological pathway for synthesizing eicosanoids. This pathway involves several steps where arachidonic acid releases from cell membrane phospholipids and converts to intermediate leukotriene A4 by 5-lipoxygenase. LTA4H then converts leukotriene A4 into leukotriene B4 which orchestrates inflammatory responses. LTA4H connects to proteins like 5-lipoxygenase which also acts within this cascade.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
Complementary Products
Primary Antibodies
AB133512
Anti-Leukotriene A4 hydrolase/LTA4H antibody [EPR5713]
RabMAb|Recombinant|KO Validated
![Western blot - Anti-Leukotriene A4 hydrolase/LTA4H antibody [EPR5713] (AB133512)](https://content.abcam.com/products/images/leukotriene-a4-hydrolase-lta4h-antibody-epr5713-ab133512--western-blot-img663.jpg)
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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