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AB261704

Human NT5E knockout A-431 cell lysate

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NT5E KO cell lysate available now. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control included. Knockout achieved by CRISPR/Cas9; X = 5 bp deletion, 2 bp deletion; Frameshift = 99.2%.

View Alternative Names

5' nucleotidase (CD73), 5' nucleotidase precursor, 5' nucleotidase, ecto, 5' nucleotidase, ecto (CD73), 5'-NT, 5'-nucleotidase, 5NTD_HUMAN, CD73, CD73 antigen, E5NT, Ecto-5'-nucleotidase, NT, NT5, NT5E, Purine 5 Prime Nucleotidase, eN, eNT

3 Images
Western blot - Human NT5E knockout A-431 cell lysate (AB261704)
  • WB

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Western blot - Human NT5E knockout A-431 cell lysate (AB261704)

Lane 1 : Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate (20 ug)
Lane 2 : NT5E knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate (20 ug)
Lane 3 : A-375 (Human malignant melanoma cell line) whole cell lysate
Lanes 1 - 3 : Merged signal (red and green). Green - ab202122 observed at 63 kDa. Red - loading control, ab181602, observed at 37 kDa.
ab202122 was shown to recognize NT5E in wild-type A-431 cells as signal was lost at the expected MW in NT5E knockout cell line ab261895 (knockout cell lysate ab261704). Additional cross-reactive bands were observed in the wild-type and knockout samples. Wild-type and NT5E knockout samples were subjected to SDS-PAGE. ab202122 and ab181602 (Rabbit anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-CD73 antibody [4G6E3] (<a href='/en-us/products/primary-antibodies/cd73-antibody-4g6e3-ab202122'>ab202122</a>) at 1/1000 dilution

Lane 1:

Wild-type A-431 whole cell lysate at 20 µg

Lane 2:

NT5E knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

Western blot - Human NT5E knockout A-431 cell line (<a href='/en-us/products/cell-lines/human-nt5e-knockout-a-431-cell-line-ab261895'>ab261895</a>)

Lane 3:

A375 (Human malignant melanoma cell line) whole cell lysate at 20 µg

Predicted band size: 63 kDa

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Western blot - Human NT5E knockout A-431 cell lysate (AB261704)
  • WB

Lab

Western blot - Human NT5E knockout A-431 cell lysate (AB261704)

Lane 1 : Wild-type (Human epidermoid carcinoma cell line) whole cell lysate (20 ug)
Lane 2 : NT5E knockout (Human epidermoid carcinoma cell line) whole cell lysate (20 ug)
Lane 3 : A-375 (Human malignant melanoma cell line) whole cell lysate
Lanes 1 - 3 : Merged signal (red and green). Green - ab133582 observed at 63 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab133582 was shown to recognize NT5E in wild-type A-431 cells as signal was lost at the expected MW in NT5E knockout cell line ab261895 (knockout cell lysate ab261704). Additional cross-reactive bands were observed in the wild-type and knockout samples. Wild-type and NT5E knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% milk. ab133582 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

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-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

NT5E knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

Western blot - Human NT5E knockout A-431 cell line (<a href='/en-us/products/cell-lines/human-nt5e-knockout-a-431-cell-line-ab261895'>ab261895</a>)

Lane 3:

A-375 (Human malignant melanoma cell line) whole cell lysate at 20 µg

Predicted band size: 105 kDa,143 kDa,24 kDa,35 kDa,41 kDa,44 kDa,45 kDa,53 kDa,58 kDa,61 kDa,62 kDa,63 kDa,70 kDa,88 kDa

Observed band size: 104 kDa,18 kDa,55 kDa,58 kDa

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Next Generation Sequencing - Human NT5E knockout A-431 cell lysate (AB261704)
  • NGS

Lab

Next Generation Sequencing - Human NT5E knockout A-431 cell lysate (AB261704)

X = 5 bp deletion, 2 bp deletion

Key facts

Cell type

A-431

Species or organism

Human

Tissue

Skin

Knockout validation

Next Generation Sequencing,Western blot

Mutation description

Knockout achieved by CRISPR/Cas9; X = 5 bp deletion, 2 bp deletion; Frameshift = 99.2%

Disease

Epidermoid 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 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
NT5E
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Next Generation 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.

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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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)

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Assay Kits

AB254513

Human CD73 ELISA Kit

Recombinant|SimpleStep

Sandwich ELISA - Human CD73 ELISA Kit (AB254513)

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Proteins & Peptides

AB310790

Recombinant Mouse CD73 protein (His tag)

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

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Product promise

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For full details, please see our Terms & Conditions

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