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AB286746

Human LATS2 knockout A549 cell line

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LATS2 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided.

View Alternative Names

FLJ13161, KPM, Kinase phosphorylated during mitosis protein, LATS large tumor suppressor Drosophila homolog 2, LATS large tumor suppressor homolog 2, LATS2_HUMAN, Large tumor suppressor homolog 2, Large tumor suppressor homolog 2 Drosophila, Large tumor suppressor kinase 2, Serine/threonine kinase kpm, Serine/threonine-protein kinase LATS2, Serine/threonine-protein kinase kpm, Warts-like kinase

2 Images
Western blot - Human LATS2 knockout A549 cell line (AB286746)
  • WB

Lab

Western blot - Human LATS2 knockout A549 cell line (AB286746)

Western blot : Anti-LATS2 antibody [EPR23126-488] (ab243657) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab243657 was shown to bind specifically to LATS2. A band was observed at 117-171 kDa in wild-type A549 cell lysates with no signal observed at this size in LATS2 knockout cell line. To generate this image, wild-type and LATS2 knockout A549 cell lysates were analysed. First, 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 IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-LATS2 antibody [EPR23126-488] (<a href='/en-us/products/primary-antibodies/lats2-antibody-epr23126-488-ab243657'>ab243657</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

Western blot - Human LATS2 knockout A549 cell line (ab286746)

Lane 2:

LATS2 knockout A549 cell lysate at 20 µg

Lane 3:

MCF7 cell lysate at 20 µg

Lane 4:

Raji cell lysate at 20 µg

Secondary

Lanes 1 - 4:

Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution

Lanes 1 - 4:

Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 117-171 kDa

false

Next Generation Sequencing - Human LATS2 knockout A549 cell line (AB286746)
  • NGS

Supplier Data

Next Generation Sequencing - Human LATS2 knockout A549 cell line (AB286746)

88 bp deletion after Tyr531

Key facts

Cell type

A549

Species or organism

Human

Tissue

Lung

Form

Liquid

form

Knockout validation

Next Generation Sequencing,Western blot

Disease

Carcinoma

Reactivity data

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

Product details

Although we aim to provide customers with a homozygous clone, feasibility will be dependent on the biology of the protein. Should only heterozygous edits be achieved, you will be notified of the outcome and be asked to confirm whether the cell line is acceptable. All clones will be accompanied with DNA sequencing data, and the mutation description.

Recommended control: Human wild-type A549 cell line (ab288558). 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.

Properties and storage information

Gene name
LATS2
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

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.
  • Do not allow the cell density to exceed 7x104 cells/cm2.
Culture medium

F-12K + 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.

LATS2 also known as Large Tumor Suppressor kinase 2 is a serine/threonine-protein kinase with a molecular mass of approximately 121 kDa. It functions as a component of the Hippo signaling pathway where it helps regulate cell proliferation apoptosis and organ size. LATS2 shows expression in various tissues with notable levels in the lung heart and liver. Its role in these tissues makes it an important target for research in cancer and developmental biology.
Biological function summary

LATS2 plays an important role in cell cycle regulation and tumor suppression. LATS2 forms part of the core kinase complex within the Hippo pathway working closely with other proteins like MST1/2 kinases and Scaffold proteins SAV1. Through phosphorylation LATS2 can inhibit the activity of YAP and TAZ transcription co-activators promoting cell contact inhibition and proper tissue architecture. Its activity is important for limiting excessive cell growth and ensuring accurate cell division.

Pathways

LATS2 integrates into the Hippo signaling pathway and influences the PI3K-AKT signaling pathway. Within the Hippo pathway LATS2 interacts with MST1/2 forming an upstream kinase complex activating through phosphorylation of downstream transcription co-activators YAP and TAZ. These interactions result in the transcriptional regulation of genes controlling cell growth and apoptosis. Overlap with the PI3K-AKT pathway links LATS2 to cellular processes related to growth migration and survival providing a critical nexus that communicates cell status and environmental cues.

LATS2 has associations with both cancer and cardiac hypertrophy. In cancer dysregulation of LATS2 activity can result in aberrant cell growth and tumor development often noted in different types of carcinomas where it can act as a tumor suppressor. Mutations or reduced expression of LATS2 can correlate with increased cancer progression implying a need for comprehensive understanding of its regulation and function. Additionally in cardiac hypertrophy LATS2 contributes to cardiac cell size regulation and its imbalance can connect to abnormal heart muscle thickening. In these contexts LATS2 interaction with proteins such as YAP and TAZ provides an important point of interest for therapeutic interventions.

Quality control

STR analysis

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

Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Adherent

Gender

Male

Product protocols

Product promise

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