Name: Anti-NNMT antibody [OTI3D8]
SKU: ab119758
Rating: 5 (2 reviews)

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Western blot - Anti-NNMT antibody [OTI3D8] (ab119758)

Key features and details

Mouse monoclonal [OTI3D8] to NNMT
Suitable for: WB, IHC-P, ICC/IF, Flow Cyt (Intra)
Knockout validated
Reacts with: Mouse, Human, Recombinant fragment
Isotype: IgG2b

Product name

Anti-NNMT antibody [OTI3D8]
See all NNMT primary antibodies
Description

Mouse monoclonal [OTI3D8] to NNMT
Host species

Mouse
Tested applications

Suitable for: WB, IHC-P, ICC/IF, Flow Cyt (Intra)more details
Species reactivity

Reacts with: Mouse, Human, Recombinant fragment
Immunogen

Recombinant full length Human NNMT, produced in HEK293T cells (NP_006160).
Positive control
- WB: HeLa, A549, HepG2 and SVT2 cell lysates; HEK293T cell lysate transfected with pCMV6-ENTRY NNMT cDNA. IHC-P: Human Liver, Pancreas, Thyroid carcinoma, Endometrium, Endometrium adenocarcinoma and Prostate tissues. ICC/IF: COS7 cells transiently transfected by pCMV6-ENTRY NNMT. Flow Cyt (Intra): Jurkat cells.
General notes

The clone number has been updated from 3D8 to OTI3D8, both clone numbers name the same clone.

This product was changed from ascites to tissue culture supernatant on 29^th May 2018. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

Form

Liquid
Storage instructions

Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid repeated freeze / thaw cycles.
Storage buffer

pH: 7.30
Preservative: 0.02% Sodium azide
Constituents: 48% PBS, 1% BSA, 50% Glycerol
Concentration information loading...
Purity

Protein A purified
Purification notes

ab119758 is purified from TCS by affinity chromatography.
Clonality

Monoclonal
Clone number

OTI3D8
Isotype

IgG2b
Research areas

Compatible Secondaries
- Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
- Goat Anti-Mouse IgG H&L (HRP) (ab205719)
Isotype control
- Mouse IgG2b, kappa monoclonal [7E10G10] - Isotype Control (ab170192)
KO cell lines
- Human NNMT knockout HeLa cell line (ab265700)
KO cell lysates
- Human NNMT knockout HeLa cell lysate (ab258537)
Recombinant Protein
- Recombinant Human NNMT protein (ab198647)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab119758 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application	Abreviews	Notes
WB	(2)	1/500 - 1/2000. Predicted molecular weight: 30 kDa.
IHC-P		1/150.
ICC/IF		1/100.
Flow Cyt (Intra)		1/100. ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

Notes
WB 1/500 - 1/2000. Predicted molecular weight: 30 kDa.
IHC-P 1/150.
ICC/IF 1/100.
Flow Cyt (Intra) 1/100. ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

Function

Catalyzes the N-methylation of nicotinamide and other pyridines to form pyridinium ions. This activity is important for biotransformation of many drugs and xenobiotic compounds.
Tissue specificity

Predominantly expressed in the liver. A lower expression is seen in the kidney, lung, skeletal muscle, placenta and heart. Not detected in the brain or pancreas.
Sequence similarities

Belongs to the NNMT/PNMT/TEMT family.
Cellular localization

Cytoplasm.
Target information above from: UniProt accession P40261 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 4837 Human
- Entrez Gene: 18113 Mouse
- Omim: 600008 Human
- SwissProt: P40261 Human
- SwissProt: O55239 Mouse
- Unigene: 503911 Human
- Unigene: 623245 Human
- Unigene: 458542 Mouse
- Unigene: 8362 Mouse
see all
Alternative names
- EC 2.1.1.1 antibody
- Nicotinamide N methyltransferase antibody
- Nicotinamide N-methyltransferase antibody
- NNMT antibody
- NNMT_HUMAN antibody
see all

Western blot - Anti-NNMT antibody [OTI3D8] (ab119758)

All lanes : Anti-NNMT antibody [OTI3D8] (ab119758) at 1/500 dilution

Lane 1 : Wild-type HeLa cell lysate
Lane 2 : NNMT knockout HeLa cell lysate
Lane 3 : A549 cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) at 1/10000 dilution

Predicted band size: 30 kDa
Observed band size: 30 kDa

Lanes 1-3: Merged signal (red and green). Green - ab119758 observed at 30 kDa. Red - loading control ab181602 observed at 36 kDa.

ab119758 Anti-NNMT antibody [OTI3D8] 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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NNMT antibody [OTI3D8] (ab119758)

ab119758 at 1/150 dilution, staining NNMT in paraffin-embedded Human liver tissue by Immunohistochemistry.
Western blot - Anti-NNMT antibody [OTI3D8] (ab119758)

All lanes : Anti-NNMT antibody [OTI3D8] (ab119758) at 1/500 dilution

Lane 1 : HEK293T cell lysate transfected with pCMV6-ENTRY control cDNA
Lane 2 : HEK293T cell lysate transfected with pCMV6-ENTRY NNMT cDNA

Lysates/proteins at 5 µg per lane.

Predicted band size: 30 kDa

HEK293T cell lysates were generated from transient transfection of the cDNA clone (RC200641)
Western blot - Anti-NNMT antibody [OTI3D8] (ab119758)

All lanes : Anti-NNMT antibody [OTI3D8] (ab119758) at 1/500 dilution

Lane 1 : HepG2 cell extracts
Lane 2 : HeLa cell extracts
Lane 3 : SVT2 cell extracts
Lane 4 : A549 cell extracts
Lane 5 : COS7 cell extracts
Lane 6 : Jurkat cell extracts
Lane 7 : MDCK cell extracts
Lane 8 : PC12 cell extracts
Lane 9 : MCF7 cell extracts

Lysates/proteins at 35 µg per lane.

Predicted band size: 30 kDa

HEK293T cell lysates were generated from transient transfection of the cDNA clone (RC200641)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NNMT antibody [OTI3D8] (ab119758)

ab119758 at 1/150 dilution, staining NNMT in paraffin-embedded Human pancreas tissue by Immunohistochemistry.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NNMT antibody [OTI3D8] (ab119758)

ab119758 at 1/150 dilution, staining NNMT in paraffin-embedded Human thyroid carcinoma tissue by Immunohistochemistry.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NNMT antibody [OTI3D8] (ab119758)

ab119758 at 1/150 dilution, staining NNMT in paraffin-embedded Human endometrium tissue by Immunohistochemistry.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NNMT antibody [OTI3D8] (ab119758)

ab119758 at 1/150 dilution, staining NNMT in paraffin-embedded Human endometrium adenocarcinoma tissue by Immunohistochemistry.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NNMT antibody [OTI3D8] (ab119758)

ab119758 at 1/150 dilution, staining NNMT in paraffin-embedded Human prostate tissue by Immunohistochemistry.
Immunocytochemistry/ Immunofluorescence - Anti-NNMT antibody [OTI3D8] (ab119758)

ab119758, at 1/100 dilution staining NNMT, in COS7 cells transiently transfected by pCMV6-ENTRY NNMT, by Immunofluorescence
Flow Cytometry (Intracellular) - Anti-NNMT antibody [OTI3D8] (ab119758)

ab119758 at 1/100 dilution staining NNMT in Jurkat cells by Flow Cytometry (Intracellular) (Red) compared to a nonspecific negative control antibody (Blue).

Flow cytometry protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

Download

Publishing research using ab119758? Please let us know so that we can cite the reference in this datasheet.

ab119758 has been referenced in 8 publications.

Cao Q et al. Long Non-coding RNA ST8SIA6-AS1 Promotes Lung Adenocarcinoma Progression Through Sponging miR-125a-3p. Front Genet 11:597795 (2020). PubMed: 33363573
Okuyama T et al. Linagliptin Ameliorates Hepatic Steatosis via Non-Canonical Mechanisms in Mice Treated with a Dual Inhibitor of Insulin Receptor and IGF-1 Receptor. Int J Mol Sci 21:N/A (2020). PubMed: 33105604
Policarpo RL et al. High-Affinity Alkynyl Bisubstrate Inhibitors of Nicotinamide N-Methyltransferase (NNMT). J Med Chem 62:9837-9873 (2019). PubMed: 31589440
Kanakkanthara A et al. BRCA1 Deficiency Upregulates NNMT, Which Reprograms Metabolism and Sensitizes Ovarian Cancer Cells to Mitochondrial Metabolic Targeting Agents. Cancer Res 79:5920-5929 (2019). PubMed: 31619387
Neelakantan H et al. Small molecule nicotinamide N-methyltransferase inhibitor activates senescent muscle stem cells and improves regenerative capacity of aged skeletal muscle. Biochem Pharmacol 163:481-492 (2019). PubMed: 30753815
Liang L et al. Nicotinamide N-methyltransferase promotes epithelial-mesenchymal transition in gastric cancer cells by activating transforming growth factor-ß1 expression. Oncol Lett 15:4592-4598 (2018). PubMed: 29541230
Ishibashi K et al. Deregulation of Nicotinamide N-Methyltransferase and Gap Junction Protein Alpha-1 Causes Metastasis in Adenoid Cystic Carcinoma. Anticancer Res 38:187-197 (2018). PubMed: 29277772
Mobley A et al. Aurora Kinase A is a Biomarker for Bladder Cancer Detection and Contributes to its Aggressive Behavior. Sci Rep 7:40714 (2017). PubMed: 28102366

Customer reviews and Q&As

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1-2 of 2 Abreviews

Application

Western blot

Sample

Human Cell lysate - whole cell (RPE)

Gel Running Conditions

Reduced Denaturing (4-20%)

Loading amount

20 µg

Specification

RPE

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: rt°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Rayne Lim

Verified customer

Submitted Aug 08 2020

Application

Western blot

Sample

Human Cell lysate - whole cell (U87 and U373 Human glioblastoma cell lines)

Loading amount

8000 cells

Specification

U87 and U373 Human glioblastoma cell lines

Gel Running Conditions

Reduced Denaturing

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Dr. Desire Ngoga

Verified customer

Submitted Jun 26 2012

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Key features and details

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Purification notes

Clonality

Clone number

Isotype

Research areas

Associated products

Compatible Secondaries

Isotype control

KO cell lines

KO cell lysates

Recombinant Protein

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Sequence similarities

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

References (8)

Customer reviews and Q&As

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