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AB258034

Human LTA4H (Leukotriene A4 hydrolase) knockout HEK-293T cell lysate

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LTA4H KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Homozygous: 10 bp deletion in exon 2.

View Alternative Names

LTA4, Leukotriene A(4) hydrolase, Leukotriene A-4 hydrolase, Tripeptide aminopeptidase LTA4H, LTA-4 hydrolase, LTA4H

3 Images
Western blot - Human LTA4H (Leukotriene A4 hydrolase) knockout HEK-293T cell lysate (AB258034)
  • WB

Lab

Western blot - Human LTA4H (Leukotriene A4 hydrolase) knockout HEK-293T cell lysate (AB258034)

Lane 1 : Wild-type HEK293T cell lysate (20 ug)
Lane 2 : LTA4H knockout HEK293T cell lysate (20 ug)
Lane 3 : T-47D cell lysate (20 ug)
Lane 4 : A549 cell lysate (20 ug)

ab109434 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 4oC 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 (<a href='/en-us/products/cell-lines/human-lta4h-leukotriene-a4-hydrolase-knockout-hek-293t-cell-line-ab266467'>ab266467</a>)

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

Western blot - Human LTA4H (Leukotriene A4 hydrolase) knockout HEK-293T cell lysate (AB258034)
  • WB

Lab

Western blot - Human LTA4H (Leukotriene A4 hydrolase) knockout HEK-293T cell lysate (AB258034)

Lane 1 : Wild-type HEK293T cell lysate (20 ug)
Lane 2 : LTA4H knockout HEK293T cell lysate (20 ug)
Lane 3 : T-47D cell lysate (20 ug)
Lane 4 : A549 cell lysate (20 ug)

ab133512 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 4oC 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 (<a href='/en-us/products/cell-lines/human-lta4h-leukotriene-a4-hydrolase-knockout-hek-293t-cell-line-ab266467'>ab266467</a>)

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 Sequencing - Human LTA4H (Leukotriene A4 hydrolase) knockout HEK-293T cell lysate (AB258034)
  • Sanger seq

Unknown

Sanger Sequencing - Human LTA4H (Leukotriene A4 hydrolase) knockout HEK-293T cell lysate (AB258034)

Homozygous : 10 bp deletion in exon 2

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 10 bp deletion in exon 2.

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Complementary Products

Primary Antibodies

AB133512

Anti-Leukotriene A4 hydrolase/LTA4H antibody [EPR5713]

RabMAb|Recombinant|KO Validated

Flow Cytometry (Intracellular) - Anti-Leukotriene A4 hydrolase/LTA4H antibody [EPR5713] (AB133512)

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Recombinant human VEGF 165A protein (Active)

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AB288799

Recombinant Human VCAM1 protein (Active)

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Functional Studies - Recombinant Human VCAM1 protein (Active) (AB288799)

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Proteins & Peptides

AB201716

Recombinant Human Leukotriene A4 hydrolase/LTA4H protein (His tag N-Terminus)

SDS-PAGE - Recombinant Human Leukotriene A4 hydrolase/LTA4H protein (His tag N-Terminus) (AB201716)

  • 0
  • 0
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Reactivity data

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Product details

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.

User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

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.

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What's included?

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Properties and storage information

Gene name
LTA4H
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Zygosity
Homozygous
Shipped at conditions
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

Leukotriene A4 hydrolase often called LTA4H is an enzyme that converts leukotriene A4 into leukotriene B4 an important mediator in inflammation. It has a molecular mass of approximately 69 kDa and functions as a zinc metalloprotease. Expression of LTA4H is widespread detected in various tissues including the lung liver and kidney. This enzyme plays an essential role in modulating inflammatory responses due to its ability to catalyze the transformation of leukotriene A4.
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.

Leukotriene A4 hydrolase associates with inflammatory diseases such as asthma and inflammatory bowel disease. In asthma the overproduction of leukotriene B4 can exacerbate airway inflammation and hyperresponsiveness. Here leukotrienes interact with receptors on effector cells which contributes to the pathology of asthma. Inflammatory bowel disease showcases a similar link where increased levels of leukotriene B4 potentiate intestinal inflammation. Effective management of these conditions may involve targeting leukotriene pathways specifically inhibiting the activity of LTA4H to ameliorate symptoms.
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Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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Product protocols

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Product promise

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For full details, please see our Terms & Conditions

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