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AB266195

Human LYPLA1 (Lysophospholipase 1/LPL-I) knockout HEK-293T cell line

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LYPLA1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 2 bp deletion in exon 1. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.

View Alternative Names

APT-1, Acyl-protein thioesterase 1, LPL-I, LPL1, LYPA1_HUMAN, LYPLA 1, LYSOPLA, LysoPLA I, Lysophospholipase 1, Lysophospholipase I, Lysophospholipid specific lysophospholipase, hAPT1

3 Images
Western blot - Human LYPLA1 (Lysophospholipase 1/LPL-I) knockout HEK-293T cell line (AB266195)
  • WB

Lab

Western blot - Human LYPLA1 (Lysophospholipase 1/LPL-I) knockout HEK-293T cell line (AB266195)

Lanes 1- 2 : Merged signal (red and green). Green - ab91606 observed at 25 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab91606 was shown to react with Lysophospholipase 1/LPL-I in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266195 (knockout cell lysate ab257509) was used. Wild-type HEK-293T and LYPLA1 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab91606 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 5000 dilution and a 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-Lysophospholipase 1/LPL-I antibody [EPR3667] (<a href='/en-us/products/primary-antibodies/lysophospholipase-1-lpl-i-antibody-epr3667-ab91606'>ab91606</a>) at 1/5000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

LYPLA1 knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human LYPLA1 (Lysophospholipase 1/LPL-I) knockout HEK-293T cell line (ab266195)

Predicted band size: 25 kDa

Observed band size: 25 kDa

false

Cell Culture - Human LYPLA1 (Lysophospholipase 1/LPL-I) knockout HEK-293T cell line (AB266195)
  • Cell Culture

Lab

Cell Culture - Human LYPLA1 (Lysophospholipase 1/LPL-I) knockout HEK-293T cell line (AB266195)

Representative images LYPLA1 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.

Sanger Sequencing - Human LYPLA1 (Lysophospholipase 1/LPL-I) knockout HEK-293T cell line (AB266195)
  • Sanger seq

Unknown

Sanger Sequencing - Human LYPLA1 (Lysophospholipase 1/LPL-I) knockout HEK-293T cell line (AB266195)

Homozygous : 2 bp deletion in exon1

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Form

Liquid

form

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 2 bp deletion in exon 1

Reactivity data

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Product details

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?

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Properties and storage information

Gene name
LYPLA1
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Zygosity
Homozygous
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C

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.

Lysophospholipase 1 also known as LPL-I is an enzyme involved in lipid metabolism. It catalyzes the hydrolysis of lysophospholipids into fatty acids and glycerophospholipids. The molecular mass of Lysophospholipase 1 is approximately 25 kDa. This protein is expressed in various tissues including heart liver and brain. Alternate names for this enzyme include APEH and acylaminoacyl-peptidase reflecting its ability to hydrolyze peptides and acyl groups.
Biological function summary

Lysophospholipase 1 plays a role in maintaining lipid homeostasis. It does not function as part of a larger protein complex acting independently in the regulation of lipid turnover. By breaking down lysophospholipids the enzyme helps balance the cellular lipid composition ensuring proper membrane integrity and cellular signaling. This regulation is critical in tissues that have high turnover of lipids like the brain and liver.

Pathways

Lysophospholipase 1 participates in lipid metabolism and glycerophospholipid biosynthesis pathways. The enzyme helps process intermediates in these pathways facilitating energy production and phospholipid remodeling. In terms of related proteins LPL-I functions in coordination with phospholipase A2 in lipid degradation and this relationship impacts the synthesis of bioactive lipid mediators.

Lysophospholipase 1 has been linked to metabolic disorders and neurological diseases. Altered activity of this enzyme has correlations with conditions such as metabolic syndrome where lipid imbalances occur. Moreover its dysregulation can influence neurological disorders like Alzheimer's Disease due to its impact on lipid signaling. In these contexts LPL-I interacts functionally with other enzymes like phospholipase A2 which also plays a role in lipid signaling and homeostasis.

Quality control

STR analysis

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

Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

Product protocols

Product promise

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