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AB256976

Human Lgals9 (galectin 9/Gal-9) knockout A549 cell lysate

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LGALS9 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon3.
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Western blot - Human Lgals9 (galectin 9/Gal-9) knockout A549 cell lysate (AB256976)
  • WB

Lab

Western blot - Human Lgals9 (galectin 9/Gal-9) knockout A549 cell lysate (AB256976)

Lane 1 : Wild-type A549 cell lysate (20 μg)

Lane 2 : LGALS9 knockout A549 cell lysate (20 μg)

Lane 3 : U-937 cell lysate (20 μg)

Lanes 1-4 : Merged signal (red and green). Green - ab227046 observed at 34-39 kDa. Red - loading control ab8245 observed at 36 kDa.

ab227046 Anti-galectin 9/Gal-9 antibody [EPR22214] was shown to specifically react with galectin 9/Gal-9 in wild-type A549 cells. Loss of signal was observed when knockout cell line ab266923 (knockout cell lysate ab256976) was used. Wild-type and galectin 9/Gal-9 knockout samples were subjected to SDS-PAGE.. ab227046 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 500 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-galectin 9/Gal-9 antibody [EPR22214] (<a href='/en-us/products/primary-antibodies/galectin-9-gal-9-antibody-epr22214-ab227046'>ab227046</a>) at 1/500 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

Galectin 9 knockout A549 cell lysate at 20 µg

Lane 2:

Western blot - Human Lgals9 (galectin 9/Gal-9) knockout A549 cell line (<a href='/en-us/products/cell-lines/human-lgals9-galectin-9-gal-9-knockout-a549-cell-line-ab266923'>ab266923</a>)

Lane 3:

U-937 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: 40 kDa

Observed band size: 34-39 kDa

false

Sanger Sequencing - Human Lgals9 (galectin 9/Gal-9) knockout A549 cell lysate (AB256976)
  • Sanger seq

Unknown

Sanger Sequencing - Human Lgals9 (galectin 9/Gal-9) knockout A549 cell lysate (AB256976)

Homozygous : 1 bp insertion in exon3

Key facts

Cell type

A549

Species or organism

Human

Tissue

Lung

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon3.

Disease

Carcinoma

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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, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and 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
LGALS9
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
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.

Galectin-9 also known as Gal-9 or LGALS9 is a β-galactoside-binding lectin with a molecular mass of around 40 kDa. It shows expression in various tissues including the thymus spleen and liver. The galectin-9 protein is involved in cell-cell and cell-matrix interactions where it binds to glycoproteins and glycolipids. Researchers commonly use methods like Galectin-9 ELISA to measure its levels in different biological samples.
Biological function summary

Galectins like galectin-9 play essential roles in regulating diverse immune responses. Galectin-9 acts within the immune system by modulating inflammation and apoptosis. It acts as both a pro-apoptotic factor for T cells and a protein mediator in innate immune functions. Galectin-9 proteins can form complexes with other lectins and cell surface receptors contributing to cellular signaling processes.

Pathways

Galectin-9 is integral to the immune system's functional networks. It participates in the T-cell regulation pathways and the innate immunity pathways. In these pathways it interacts with other proteins like TIM-3 which mediates T-cell apoptosis and contributes to immune tolerance. These interaction networks play important roles in maintaining the body's immune homeostasis.

Galectin-9 has associations with autoimmune diseases and cancer. Its regulatory effects on T cells connect it to autoimmune conditions where abnormal T-cell activation occurs. In cancer the alteration in galectin-9 expression levels affects tumor immunity. The interaction of galectin-9 with other proteins like TIM-3 is important in cancer progression making it a target of interest for therapeutic interventions.
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Quality control

STR analysis

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

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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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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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