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AB267013

Human NMI (N myc interactor/NMI) knockout A549 cell line

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NMI KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 2 bp deletion in exon 5 and 330 bp insertion in exon 5. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
4 Images
Western blot - Human NMI (N myc interactor/NMI) knockout A549 cell line (AB267013)
  • WB

Lab

Western blot - Human NMI (N myc interactor/NMI) knockout A549 cell line (AB267013)

Lanes 1-4 : Merged signal (red and green). Green - ab183724 observed at 39 kDa. Red - loading control ab8245 observed at 36 kDa.

ab183724 Anti-N myc interactor/NMI antibody [EPR11065(2)] was shown to specifically react with N myc interactor/NMI in wild-type A549 cells. Loss of signal was observed when knockout cell line ab267013 (knockout cell lysate ab258077) was used. Wild-type and N myc interactor/NMI knockout samples were subjected to SDS-PAGE. ab183724 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-N myc interactor/NMI antibody [EPR11065(2)] (<a href='/en-us/products/primary-antibodies/n-myc-interactor-nmi-antibody-epr110652-ab183724'>ab183724</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

NMI knockout A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

Western blot - Human NMI (N myc interactor/NMI) knockout A549 cell line (ab267013)

Lane 3:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 4:

K562 (Human chronic myelogenous leukemia lymphoblast cell line ) 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: 35 kDa

Observed band size: 39 kDa

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Cell Culture - Human NMI (N myc interactor/NMI) knockout A549 cell line (AB267013)
  • Cell Culture

Lab

Cell Culture - Human NMI (N myc interactor/NMI) knockout A549 cell line (AB267013)

Representative images NMI knockout A549 cells, low and high confluency examples (top left and right respectively) and wild-type A549 cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS M5000 microscope.

Sanger Sequencing - Human NMI (N myc interactor/NMI) knockout A549 cell line (AB267013)
  • Sanger seq

Unknown

Sanger Sequencing - Human NMI (N myc interactor/NMI) knockout A549 cell line (AB267013)

Allele-2 : 330 bp insertion in exon 5.

Sanger Sequencing - Human NMI (N myc interactor/NMI) knockout A549 cell line (AB267013)
  • Sanger seq

Unknown

Sanger Sequencing - Human NMI (N myc interactor/NMI) knockout A549 cell line (AB267013)

Allele-1 : 2 bp deletion in exon5

Key facts

Cell type

A549

Species or organism

Human

Tissue

Lung

Form

Liquid

form

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 2 bp deletion in exon 5 and 330 bp insertion in exon 5

Disease

Carcinoma

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Complementary Products

Cell Lines & Lysates

AB256500

Human MYC (c-Myc) knockout HEK-293T cell line

Advanced Validation

Immunocytochemistry/ Immunofluorescence - Human MYC (c-Myc) knockout HEK-293T cell line (AB256500)

  • 5
  • 1
Primary Antibodies

AB183724

Anti-N myc interactor/NMI antibody [EPR11065(2)]

RabMAb|Recombinant|KO Validated

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-N myc interactor/NMI antibody [EPR11065(2)] (AB183724)

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  • 0
Proteins & Peptides

AB103504

Recombinant Human N myc interactor/NMI protein

SDS-PAGE - Recombinant Human N myc interactor/NMI protein (AB103504)

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  • 0
Cell Lines & Lysates

AB258077

Human NMI (N myc interactor/NMI) knockout A549 cell lysate

Western blot - Human NMI (N myc interactor/NMI) knockout A549 cell lysate (AB258077)

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  • 0
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Reactivity data

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

We will provide viable cells that proliferate on revival.

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
NMI
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.
  • Do not allow the cell density to exceed 7x104 cells/cm2.
Culture medium

F-12K + 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.

N myc interactor often referred to as NMI interacts with members of the MYC family including N-MYC and C-MYC. Known as N-MYC and C-MYC interacting protein NMI has a molecular weight of approximately 54 kDa. NMI expresses in a variety of tissues with higher levels in brain and liver tissues. Its presence is also notable in immune cells pointing towards a complex role in cellular processes related to both developmental and immune responses.
Biological function summary

NMI collaborates with transcription factors to influence the regulation of gene expression. It plays a significant role in the formation of protein complexes that regulate transcriptional activities of oncogenes particularly through interaction with MYC and STAT proteins. NMI's ability to form complexes makes it a notable player in modulating signal transduction pathways. Its balancing act with transcription factors affects cell proliferation and differentiation critical components of its biological functionality.

Pathways

NMI has a significant impact on the JAK-STAT signaling pathway and the MYC-mediated transcription network. Within these pathways NMI modulates the activities of the MYC family of oncogenes and interacts with signal transducers and activators like STATs. These interactions contribute to the regulation of cellular processes such as growth and apoptosis. Such pathways illustrate NMI's role in maintaining cellular homeostasis and its interactive role with proteins like STAT and MYC.

NMI has been associated with cancer and autoimmune diseases. NMI's interaction with MYC oncogenes links it to various cancers particularly neuroblastoma. It also associates with autoimmune disorder mechanisms implicating its expression and regulation in immune-related pathologies. In cancer NMI's relationship with MYC affects tumor growth and progression illustrating its potential as a target for therapeutic intervention. Its involvement highlights the importance of understanding its function in disease pathways.
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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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Target data

See full target information NMI
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Product promise

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