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AB258083

Human NTPCR (Nucleoside triphosphate phosphohydrolase) knockout HEK-293T cell lysate

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NTPCR KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 29 bp deletion in exon 1 and 2 bp deletion in exon 1.

View Alternative Names

C1orf57, Chromosome 1 open reading frame 57, FLJ11383, HCR NTPase, Human cancer related NTPase, Hypothetical protein LOC84284, MGC13186, Novel DUF829 domain containing protein, Nucleoside triphosphatase C1orf57, Nucleoside triphosphatase, cancer related, Nucleoside triphosphate phosphohydrolase, OTTHUMP00000035941, OTTHUMP00000035942, Probable UPF0334 kinase like protein C1orf57, RP4 659I19.2, RP4 678E16.2

4 Images
Western blot - Human NTPCR (Nucleoside triphosphate phosphohydrolase) knockout HEK-293T cell lysate (AB258083)
  • WB

Unknown

Western blot - Human NTPCR (Nucleoside triphosphate phosphohydrolase) knockout HEK-293T cell lysate (AB258083)

Lane 1 : Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate (20 ug)
Lane 2 : NTPCR knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate (20 ug)
Lane 3 : U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) whole cell lysate (20 ug)
Lane 4 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate (20 ug)

ab182154 was shown to specifically react with Nucleoside triphosphate phosphohydrolase in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266397 (knockout cell lysate ab258083) was used. Wild-type and Nucleoside triphosphate phosphohydrolase knockout samples were subjected to SDS-PAGE. ab182154 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4oC at 1 in 1000 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-Nucleoside triphosphate phosphohydrolase antibody [EPR14325] (<a href='/en-us/products/primary-antibodies/nucleoside-triphosphate-phosphohydrolase-antibody-epr14325-ab182154'>ab182154</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg

Lane 2:

NTPCR knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg

Lane 2:

Western blot - Human NTPCR (Nucleoside triphosphate phosphohydrolase) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-ntpcr-nucleoside-triphosphate-phosphohydrolase-knockout-hek-293t-cell-line-ab266397'>ab266397</a>)

Lane 3:

U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) whole cell lysate at 20 µg

Lane 4:

Jurkat (Human T cell leukemia cell line from peripheral blood) whole 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: 21 kDa

Observed band size: 21 kDa

false

Western blot - Human NTPCR (Nucleoside triphosphate phosphohydrolase) knockout HEK-293T cell lysate (AB258083)
  • WB

Lab

Western blot - Human NTPCR (Nucleoside triphosphate phosphohydrolase) knockout HEK-293T cell lysate (AB258083)

Lane 1 : Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate (20 ug)
Lane 2 : NTPCR knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate (20 ug)
Lane 3 : U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) whole cell lysate (20 ug)
Lane 4 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate (20 ug)

ab182155 was shown to specifically react with Nucleoside triphosphate phosphohydrolase in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266397 (knockout cell lysate ab258083) was used. Wild-type and Nucleoside triphosphate phosphohydrolase knockout samples were subjected to SDS-PAGE. ab182155 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4oC at 1 in 1000 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-Nucleoside triphosphate phosphohydrolase antibody [EPR14325-50] - N-terminal (<a href='/en-us/products/primary-antibodies/nucleoside-triphosphate-phosphohydrolase-antibody-epr14325-50-n-terminal-ab182155'>ab182155</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg

Lane 2:

NTPCR knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg

Lane 2:

Western blot - Human NTPCR (Nucleoside triphosphate phosphohydrolase) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-ntpcr-nucleoside-triphosphate-phosphohydrolase-knockout-hek-293t-cell-line-ab266397'>ab266397</a>)

Lane 3:

U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) whole cell lysate at 20 µg

Lane 4:

Jurkat (Human T cell leukemia cell line from peripheral blood) whole 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: 21 kDa

Observed band size: 21 kDa

false

Sanger Sequencing - Human NTPCR (Nucleoside triphosphate phosphohydrolase) knockout HEK-293T cell lysate (AB258083)
  • Sanger seq

Unknown

Sanger Sequencing - Human NTPCR (Nucleoside triphosphate phosphohydrolase) knockout HEK-293T cell lysate (AB258083)

Allele-2 : 2 bp deletion in exon 1

Sanger Sequencing - Human NTPCR (Nucleoside triphosphate phosphohydrolase) knockout HEK-293T cell lysate (AB258083)
  • Sanger seq

Unknown

Sanger Sequencing - Human NTPCR (Nucleoside triphosphate phosphohydrolase) knockout HEK-293T cell lysate (AB258083)

Allele-1 : 29 bp deletion in exon 1

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 29 bp deletion in exon 1 and 2 bp deletion in exon 1.

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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?

{ "values": { "1Kit": { "sellingSize": "1 Kit", "publicAssetCode":"ab258083-1Kit", "assetComponentDetails": [ { "size":"1 x 100 µg", "name":"Human NTPCR knockout HEK293T cell lysate", "number":"AB258083-CMP01", "productcode":"" }, { "size":"1 x 100 µg", "name":"Human wild-type HEK293T cell lysate", "number":"AB258083-CMP02", "productcode":"" } ] } } }
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Properties and storage information

Gene name
NTPCR
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.

Nucleoside triphosphate phosphohydrolase often called NTPase is an enzyme responsible for the hydrolysis of nucleoside triphosphates. It plays an important role in converting nucleoside triphosphates to nucleoside diphosphates with the release of inorganic phosphate. This process is important as it regulates the availability of nucleotides for various biological functions. NTPase typically with a mass of approximately 70 kDa is expressed in various tissues but shows high activity in tissues with rapid cell division. It is particularly studied in cellular lines such as HEK293T where it assists in the investigation of cellular mechanisms.
Biological function summary

This enzyme participates in critical cellular processes by managing the concentration of nucleoside triphosphates. Its activity influences DNA and RNA synthesis signal transduction and energy transfer. NTPase does not generally form complexes with other proteins but may interact with other molecules transiently during its catalytic action. This transient nature allows it to act efficiently where nucleoside triphosphate function is necessary.

Pathways

NTPase influences multiple biochemical pathways important for cell viability and function. It plays a role in the nucleotide metabolism pathway which is essential for DNA and RNA biosynthesis. Moreover its involvement in the energy transfer pathway ties it to proteins such as myosin and kinases which require energy from ATP hydrolysis to function properly. These interactions underline its importance in cellular physiology.

Disruptions in NTPase activity have been linked to mitochondrial disorders and certain cancers. Abnormal levels of nucleoside triphosphates caused by malfunctioning NTPase can lead to mitochondrial dysfunction contributing to disorders like mitochondrial myopathy. In cancer altered nucleotide metabolism may support uncontrolled cell proliferation. NTPase could connect with proteins such as p53 or oncogene products accumulating interest as a potential therapeutic target.
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Quality control

STR analysis

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

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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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Product protocols

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