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AB306752

Human PLA2G6 knockout U-87 MG cell line

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PLA2G6 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control available. Knockout. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
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Next Generation Sequencing - Human PLA2G6 knockout U-87 MG cell line (AB306752)
  • NGS

Lab

Next Generation Sequencing - Human PLA2G6 knockout U-87 MG cell line (AB306752)

22 bp deletion (allele 1), 1 bp and 8 bp deletion (allele 2) in exon 1, CCDS13967.1

Western blot - Human PLA2G6 knockout U-87 MG cell line (AB306752)
  • WB

Lab

Western blot - Human PLA2G6 knockout U-87 MG cell line (AB306752)

Western blot : Rabbit Monoclonal[EPR23994-103] to Calcium-independent Phospholipase A2/PLA2G6 ab259950 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH (ab8245) loading control staining at 1/20,000 dilution, shown in magenta. A band was observed at 76 kDa in Wild-type U-87 MG cell lysates with no signal observed at this size in PLA2G6 knockout U-87 MG cell line. To generate this image, 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 800CW & Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-Calcium-independent Phospholipase A2/PLA2G6 antibody [EPR23994-103] (<a href='/en-us/products/primary-antibodies/calcium-independent-phospholipase-a2-pla2g6-antibody-epr23994-103-ab259950'>ab259950</a>) at 1/1000 dilution

Lane 1:

Wild-type U-87 MG at 20 µg

Lane 2:

Western blot - Human PLA2G6 knockout U-87 MG cell line (ab306752) at 20 µg

Lane 3:

LNCaP at 20 µg

Lane 4:

Human Brain at 20 µg

Lane 5:

PC-3 at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 90 kDa

Observed band size: 76 kDa

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Key facts

Cell type

U-87 MG

Species or organism

Human

Tissue

Brain

Form

Liquid

form

Knockout validation

Next Generation Sequencing,Western blot

Mutation description

Knockout.

Disease

Glioblastoma

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Complementary Products

Primary Antibodies

AB259950

Anti-Calcium-independent Phospholipase A2/PLA2G6 antibody [EPR23994-103]

BOND RX™ Validated|KO Validated|RabMAb|Recombinant

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calcium-independent Phospholipase A2/PLA2G6 antibody [EPR23994-103] (AB259950)

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "NGS": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
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Product details

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
PLA2G6
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Next Generation Sequencing, Western blot
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C
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Handling procedures

Initial handling 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.
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

EMEM + 10% FBS

Cryopreservation medium

Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

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Supplementary information

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

The protein Calcium-independent Phospholipase A2 also known as iPLA2 or PLA2G6 is an important enzyme involved in phospholipid metabolism. This enzyme exhibits a catalytic function that does not require calcium for activation which distinguishes it from other members of the phospholipase A2 family. It possesses a molecular weight of approximately 85 kDa. This protein mainly expresses in cytosolic compartments within several tissues including the heart brain and pancreas playing a diverse role in cellular activities.
Biological function summary

The protein catalyzes the hydrolysis of ester bonds within phospholipids releasing free fatty acids and lysophospholipids. This enzyme does not operate as part of a larger complex but functions independently within the cell membrane and metabolic pathways. By regulating phospholipid turnover and remodeling the protein plays a vital role in membrane homeostasis and signaling. This remodeling contributes to cell survival and apoptosis depending on cellular needs and conditions.

Pathways

This enzyme interacts with lipid signaling and metabolism in central biological processes. It plays a significant role in the glycerophospholipid and arachidonic acid pathways. In these pathways iPLA2 is closely related to cyclooxygenase and lipoxygenase enzymes which convert arachidonic acid into prostaglandins and leukotrienes leading to the mediation of inflammation and other signaling pathways. This interaction emphasizes the enzyme's importance in both normal physiology and pathophysiological conditions.

Mutations or altered activities of iPLA2 have associations with neurodegenerative disorders such as Parkinson’s disease and infantile neuroaxonal dystrophy. In these disorders mitochondrial dysfunction often linked with proteins like alpha-synuclein becomes prevalent demonstrating the enzyme's influence on apoptosis and oxidative stress. Furthermore iPLA2 mutations may lead to cardiomyopathy where its function overlaps with creatine kinase and enzymes involved in energy metabolism indicating a broader impact on cardiac health.
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Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Adherent

Gender

Male

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Product protocols

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Target data

See full target information PLA2G6
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Product promise

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