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AB266215

Human NME1 (NM23A) knockout HEK-293T cell line

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NME1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 2 bp deletion in exon 2 and 5 bp deletion in exon 2.

View Alternative Names

AWD, AWD, drosophila, homolog of, GAAD, GZMA activated DNase, Granzyme A-activated DNase, Metastasis inhibition factor NM23, NB, NBS, NDKA_HUMAN, NDP kinase A, NDPK-A, NM23, NM23 long variant, included, NM23-M1, NM23H1B, included, NME/NM23 nucleoside diphosphate kinase 1, NME1-NME2 spliced read-through transcript, included, Nme1, Non-metastatic cells 1, protein (NM23A) expressed in, Nonmetastatic cells 1, protein expressed in, Nonmetastatic protein 23, Nonmetastatic protein 23, homolog 1, Nucleoside diphosphate kinase A, Tumor metastatic process-associated protein, nm23-H1

5 Images
Western blot - Human NME1 (NM23A) knockout HEK-293T cell line (AB266215)
  • WB

Lab

Western blot - Human NME1 (NM23A) knockout HEK-293T cell line (AB266215)

Lanes 1-4 : Merged signal (red and green). Green - ab171935 observed at 17 kDa. Red - loading control ab8245 observed at 36 kDa.

ab171935 Anti-NM23A antibody [EPR10146] was shown to specifically react with NM23A in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266215 (knockout cell lysate ab258075) was used. Wild-type and NM23A knockout samples were subjected to SDS-PAGE. ab171935 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° 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-NM23A antibody [EPR10146] (<a href='/en-us/products/primary-antibodies/nm23a-antibody-epr10146-ab171935'>ab171935</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

NME1 knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human NME1 (NM23A) knockout HEK-293T cell line (ab266215)

Lane 3:

MCF7 cell lysate at 20 µg

Lane 4:

Jurkat 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: 17 kDa

Observed band size: 17 kDa

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Western blot - Human NME1 (NM23A) knockout HEK-293T cell line (AB266215)
  • WB

Lab

Western blot - Human NME1 (NM23A) knockout HEK-293T cell line (AB266215)

Lanes 1-4 : Merged signal (red and green). Green - ab92327 observed at 17 kDa. Red - loading control ab8245 observed at 36 kDa.

ab92327 Anti-NM23A antibody [EPR3036] was shown to specifically react with NM23A in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266215 (knockout cell lysate ab258075) was used. Wild-type and NM23A knockout samples were subjected to SDS-PAGE. ab92327 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C 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-NM23A antibody [EPR3036] (<a href='/en-us/products/primary-antibodies/nm23a-antibody-epr3036-ab92327'>ab92327</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

NME1 knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human NME1 (NM23A) knockout HEK-293T cell line (ab266215)

Lane 3:

MCF7 cell lysate at 20 µg

Lane 4:

Jurkat 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: 17 kDa

Observed band size: 17 kDa

false

Sanger Sequencing - Human NME1 (NM23A) knockout HEK-293T cell line (AB266215)
  • Sanger seq

Unknown

Sanger Sequencing - Human NME1 (NM23A) knockout HEK-293T cell line (AB266215)

Allele-1 : 5 bp deletion in exon2

Sanger Sequencing - Human NME1 (NM23A) knockout HEK-293T cell line (AB266215)
  • Sanger seq

Unknown

Sanger Sequencing - Human NME1 (NM23A) knockout HEK-293T cell line (AB266215)

Allele-2 : 2 bp deletion in exon 2.

Cell Culture - Human NME1 (NM23A) knockout HEK-293T cell line (AB266215)
  • Cell Culture

Unknown

Cell Culture - Human NME1 (NM23A) knockout HEK-293T cell line (AB266215)

Representative images of NME1 knockout HEK293T cells, low and high confluency examples (top left and right respectively) and wild-type HEK293T cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using an EVOS M5000 microscope.

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Form

Liquid

form

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 2 bp deletion in exon 2 and 5 bp deletion in exon 2

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Reactivity data

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Product details

Recommended control: Human wild-type HEK293T cell line (ab255449). 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.

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What's included?

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Properties and storage information

Gene name
NME1
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C
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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.
Culture medium

DMEM (High Glucose) + 10% FBS

Cryopreservation medium

Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

NM23A also known as NME1 (non-metastatic cells 1 protein) is a nucleoside diphosphate kinase A. NM23A weighs approximately 17 kDa. This protein is expressed in a variety of tissues with high levels in liver brain and endocrine tissues. By exchanging phosphate groups NM23A is involved in energy transfer in cells. It stabilizes proteins by forming hexameric complexes which are pivotal for its function.
Biological function summary

The NME1 protein plays a significant role in cell proliferation and differentiation. It acts as a marker for the metastatic potential of some tumors where lower expression links to increased metastatic abilities in tumor cells. This protein sometimes works as part of a larger multiprotein complex which influences its functions in cellular processes. It is known for modulating microtubule assembly which affects cell structure and mechanics.

Pathways

NM23A is an important player in signal transduction and DNA repair pathways. It is essential in the regulation of the Ras-MAPK pathway which controls cell growth and differentiation. NM23A interacts with key regulatory proteins such as c-Myc and APC to uphold cellular activities. The protein's influence in these pathways indicates its role in sustaining normal cellular functions and responses.

NM23A relates closely to cancer particularly in metastatic processes and tumor progression. Lower levels of NM23A are associated with the progression of neuroblastoma and breast cancer. In breast cancer NM23A links to the tumor suppressor gene p53 affecting its regulatory functions. Research continues into how NM23A can serve as a useful biomarker for prognosis and therapy in cancer treatment.
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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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Target data

See full target information NME1
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