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AB282943

Human NT5E knockout A375 cell lysate

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NT5E KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Homozygous: 121 bp deletion in exon 2.
3 Images
Western blot - Human NT5E knockout A375 cell lysate (AB282943)
  • WB

Lab

Western blot - Human NT5E knockout A375 cell lysate (AB282943)

Lane 1 : Wild-type A-375 cell lysate 20 μg
Lane 2 : NT5E knockout A-375 cell lysate 20 μg
Lane 3 : A431 cell lysate 20 μg
Lane 4 : HepG2 cell lysate 20 μg
False colour image of Western blot : Anti-CD73 antibody [4G6E3] staining at 1/500 dilution, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab202122 was shown to bind specifically to CD73. A band was observed at 65-70 kDa in wild-type A-375 cell lysates with no signal observed at this size in NT5E knockout cell line ab281591 (knockout cell lysate ab282943). A cross-reactive band is observed at 65 kDa in all samples. To generate this image, wild-type and NT5E knockout A-375 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-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) at 1/20000 dilution.

All lanes:

Western blot - Anti-CD73 antibody [4G6E3] (<a href='/en-us/products/primary-antibodies/cd73-antibody-4g6e3-ab202122'>ab202122</a>) at 1/500 dilution

Lane 1:

Wild-type A-375 cell lysate at 20 µg

Lane 2:

NT5E knockout A-375 cell lysate at 20 µg

Lane 2:

Western blot - Human NT5E knockout A375 cell line (<a href='/en-us/products/cell-lines/human-nt5e-knockout-a375-cell-line-ab281591'>ab281591</a>)

Lane 3:

A431 cell lysate at 20 µg

Lane 4:

HepG2 cell lysate at 20 µg

Predicted band size: 63 kDa

Observed band size: 65 kDa,65-70 kDa

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Western blot - Human NT5E knockout A375 cell lysate (AB282943)
  • WB

Lab

Western blot - Human NT5E knockout A375 cell lysate (AB282943)

All lanes:

Western blot - Anti-CD73 antibody [EPR6114] (<a href='/en-us/products/primary-antibodies/cd73-antibody-epr6114-ab133582'>ab133582</a>) at 1/1000 dilution

Lane 1:

Wild-type A-375 cell lysate at 20 µg

Lane 2:

NT5E knockout A-375 cell lysate at 20 µg

Lane 3:

A431 cell lysate at 20 µg

Lane 4:

HepG2 cell lysate at 20 µg

Secondary

Lanes 1 - 4:

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 1/20000 dilution

Lanes 1 - 4:

Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/20000 dilution

Observed band size: 70 kDa

false

Sanger Sequencing - Human NT5E knockout A375 cell lysate (AB282943)
  • Sanger seq

Lab

Sanger Sequencing - Human NT5E knockout A375 cell lysate (AB282943)

121 bp deletion in exon 2

Key facts

Cell type

A-375

Species or organism

Human

Tissue

Skin

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 121 bp deletion in exon 2

Disease

Melanoma

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Complementary Products

Primary Antibodies

AB133582

Anti-CD73 antibody [EPR6114]

RabMAb|Recombinant|KO Validated

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD73 antibody [EPR6114] (AB133582)

  • 3
  • 2
Assay Kits

AB254513

Human CD73 ELISA Kit

Recombinant|SimpleStep

Sandwich ELISA - Human CD73 ELISA Kit (AB254513)

  • 0
  • 0
Proteins & Peptides

AB310790

Recombinant Mouse CD73 protein (His tag)

SDS-PAGE - Recombinant Mouse CD73 protein (His tag) (AB310790)

  • 0
  • 0
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Reactivity data

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

{ "values": { "1Kit": { "sellingSize": "1 Kit", "publicAssetCode":"ab282943-1Kit", "assetComponentDetails": [ { "size":"1 x 100 µg", "name":"Human NT5E knockout A375 cell lysate", "number":"AB282943-CMP01", "productcode":"" }, { "size":"1 x 100 µg", "name":"Human wild-type A375 cell lysate", "number":"AB282943-CMP02", "productcode":"" } ] } } }
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Properties and storage information

Gene name
NT5E
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.

CD73 also known as ecto-5'-nucleotidase is an enzyme that functions at the cellular level. The CD73 protein hydrolyzes extracellular nucleotides into nucleosides by removing phosphate groups. It is a 70 kDa glycosylphosphatidylinositol-anchored molecule found on the surface of various cell types including endothelial cells lymphocytes and epithelial cells. Its presence is noted in a wide range of tissues across the human body highlighting its versatile role in cellular communication and metabolism.
Biological function summary

CD73 participates in the regulation of purinergic signaling through adenosine production. It acts in cooperation with other cell surface enzymes such as CD39 forming a functional complex that handles ATP breakdown to adenosine. This enzymatic activity modulates immune responses tissue protection and inflammation control. In addition to immune-related functions CD73 supports the maintenance of vascular integrity and proper cellular adhesion which is important in different physiological and pathological contexts.

Pathways

CD73 plays an important part in the adenosinergic pathway governing the conversion of AMP into adenosine. This activity impacts the signaling and function of the adenosine receptors on immune and non-immune cells. Another important pathway is the angiogenesis process where CD73 contributes by affecting endothelial cell migration and blood vessel development. Relationships with key proteins such as adenosine A2 receptors help facilitate these critical pathway activities.

CD73's involvement with immunosuppression and tumor progression becomes significant. Elevated levels of CD73 are frequently observed in some cancers where it aids in creating a favorable tumor microenvironment through increased adenosine production. This situation assists tumor cells in evading immune detection. Additionally CD73 is linked to autoimmune diseases like multiple sclerosis where it contributes to modulated lymphocyte activity. Connections with proteins such as TGF-beta further illustrate its role in disease-related pathways creating potential targets for therapeutic interventions through the use of anti-CD73 antibodies.
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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

Female

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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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