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AB258537

Human NNMT knockout HeLa cell lysate

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NNMT KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 16 bp deletion in exon1 and 1 bp insertion in exon1.
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Western blot - Human NNMT knockout HeLa cell lysate (AB258537)
  • WB

Lab

Western blot - Human NNMT knockout HeLa cell lysate (AB258537)

Lane 1 : Wild-type HeLa cell lysate (20 ug)
Lane 2 : NNMT knockout HeLa cell lysate (20 ug)
Lane 3 : A549 cell lysate (20 ug)

ab119758 was shown to specifically react with NNMT in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265700 (knockout cell lysate ab258537) was used. Wild-type and NNMT knockout samples were subjected to SDS-PAGE. ab119758 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) were incubated at room temperature for 2.5 hours at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-NNMT antibody [OTI3D8] (<a href='/en-us/products/primary-antibodies/nnmt-antibody-oti3d8-ab119758'>ab119758</a>) at 1/500 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

NNMT knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human NNMT knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-nnmt-knockout-hela-cell-line-ab265700'>ab265700</a>)

Lane 3:

A549 cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-680rd-preadsorbed-ab216777'>ab216777</a>) at 1/10000 dilution

Predicted band size: 30 kDa

Observed band size: 30 kDa

false

Sanger Sequencing - Human NNMT knockout HeLa cell lysate (AB258537)
  • Sanger seq

Unknown

Sanger Sequencing - Human NNMT knockout HeLa cell lysate (AB258537)

Allele-2 : 1 bp insertion in exon1

Sanger Sequencing - Human NNMT knockout HeLa cell lysate (AB258537)
  • Sanger seq

Unknown

Sanger Sequencing - Human NNMT knockout HeLa cell lysate (AB258537)

Allele-1 : 16 bp deletion in exon1

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 16 bp deletion in exon1 and 1 bp insertion in exon1.

Disease

Adenocarcinoma

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Complementary Products

Primary Antibodies

AB119758

Anti-NNMT antibody [OTI3D8]

KO Validated

Western blot - Anti-NNMT antibody [OTI3D8] (AB119758)

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Proteins & Peptides

AB259412

Recombinant human VEGF 165A protein (Active)

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Mass Spectrometry - Recombinant human VEGF 165A protein (Active) (AB259412)

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Proteins & Peptides

AB180317

Recombinant Human POLR1C protein

SDS-PAGE - Recombinant Human POLR1C protein (AB180317)

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Proteins & Peptides

AB271638

Recombinant human NNMT protein (Active)

Functional Studies - Recombinant human NNMT protein (Active) (AB271638)

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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, 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
NNMT
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.

The NNMT protein also known as Nicotinamide N-methyltransferase acts by catalyzing the methylation of nicotinamide using S-adenosylmethionine as a methyl donor. This enzyme has a mass of around 29 kDa and is expressed in various tissues including liver adipose tissue and certain cancer cells. The NNMT gene provides instructions for creating the protein that plays significant roles in metabolic processes and cellular homeostasis.
Biological function summary

NNMT impacts cell energy balance and gene expression regulation by influencing the levels of nicotinamide and methionine metabolites. This enzyme does not typically form part of a complex acting more so as an individual regulator of methylation reactions affecting cellular metabolism. Researchers find interest in NNMT for its role in modulating cellular growth and proliferation forming a link to both normal cellular functions and pathological conditions like cancer.

Pathways

NNMT has a significance in NAD+ metabolism and methionine salvage pathways. These pathways are essential in maintaining redox homeostasis and proper methylation status in cells. NNMT through its enzymatic activity interacts with SAM (S-adenosyl methionine) closely relating its function to methylation processes that involve other proteins such as SIRTs (Sirtuins) and NAMPT (Nicotinamide phosphoribosyltransferase) within these metabolic pathways.

NNMT shows relevance in cancer and obesity-related complications. The protein's upregulation in cancer types like breast and colorectal cancers links it to tumor development and progression. In obesity NNMT's involvement in adipose tissue might contribute to metabolic dysregulation. The enzyme also interconnects with proteins like insulin receptors and adiponectin highlighting its potential role in metabolic diseases and making it an interesting target for therapeutic interventions.
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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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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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