重组Anti-NNMT抗体[EPR29205-77] - BSA and Azide free (ab318281)
![Western blot - Anti-NNMT antibody [EPR29205-77] - BSA and Azide free (ab318281)](https://www.abcam.cn/ps/products/318/ab318281/Images/ab318281-544359-anti-nnmt-antibody-epr29205-77-western-blot-wild-type-hela-human-cervical-adenocarcinoma-epithelial-cell-whole-cell-lysate20-ug.jpg "Western blot - Anti-NNMT antibody [EPR29205-77] - BSA and Azide free (ab318281)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR29205-77] to NNMT - BSA and Azide free
- Suitable for: IHC-P, ICC/IF, WB
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
概述
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产品名称
Anti-NNMT抗体[EPR29205-77] - BSA and Azide free
参阅全部 NNMT 一抗 -
描述
兔单克隆抗体[EPR29205-77] to NNMT - BSA and Azide free -
宿主
Rabbit -
经测试应用
适用于: IHC-P, ICC/IF, WBmore details
不适用于: Flow Cyt (Intra) -
种属反应性
与反应: Human
不与反应: Mouse, Rat -
免疫原
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: Wild-type HeLa whole cell lysate, 786-O whole cell lysate, Human liver tissue, HUVEC whole cell lysate, A549 whole cell lysate, His-tagged human NNMT recombinant protein lysates. IHC-P: Human liver, Human colon carcinoma, Human breast carcinoma, and Wild-type Hela tissues. ICC/IF: HeLa cell.
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常规说明
ab318281 is the carrier-free version of ab318280
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C. -
存储溶液
pH: 7.20
Constituent: 100% PBS -
无载体
是 -
Concentration information loading... -
纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR29205-77 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Compatible Secondaries
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KO cell lines
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KO cell lysates
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab318281于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。
| 应用 | Ab评论 | 说明 |
|---|---|---|
| IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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| ICC/IF |
Use at an assay dependent concentration.
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| WB |
Use at an assay dependent concentration.
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| 说明 |
|---|
|
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
|
ICC/IF
Use at an assay dependent concentration. |
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WB
Use at an assay dependent concentration. |
靶标
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功能
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. -
组织特异性
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. -
序列相似性
Belongs to the NNMT/PNMT/TEMT family. -
细胞定位
Cytoplasm. - Information by UniProt
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数据库链接
- Entrez Gene: 4837 Human
- Omim: 600008 Human
- SwissProt: P40261 Human
- Unigene: 503911 Human
- Unigene: 623245 Human
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别名
- EC 2.1.1.1 antibody
- Nicotinamide N methyltransferase antibody
- Nicotinamide N-methyltransferase antibody
see all
图片
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Western blot - Anti-NNMT antibody [EPR29205-77] - BSA and Azide free (ab318281)All lanes : Anti-NNMT antibody [EPR29205-77] (ab318280) at 1/1000 dilution
Lane 1 : Wild-type HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : NNMT knockout HeLa whole cell lysate
Lane 3 : 786-O (human kidney epithelial cell) whole cell lysate
Lane 4 : MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (800CW) and Goat Anti-Mouse IgG H&L (680RD) at 1/20000 dilution
Observed band size: 29 kDa why is the actual band size different from the predicted?This data was developed using ab318280, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: Intercept® (TBS) Blocking Buffer diluted with an equal volume of TBS.
The samples were run on a Bis-Tris gel under reducing conditions.
Western blot: Anti-NNMT antibody (ab318280) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta.
In Western blot, ab318280 was shown to bind specifically to NNMT. Target of interest was observed at 29 kDa in wild-type HeLa cell lysates (lane 1) with no signal observed at this size in NNMT knockout cell line (lane 2, knockout cell line ab265700 / knockout cell lysate ab258537). To generate this image, samples were first run on an SDS-PAGE gel then transferred onto an immobilon-FL PVDF membrane. Membranes were blocked in a fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4 °C. Blots were washed in TBS-T, incubated with secondary antibodies Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution for 1 h at room temperature, washed again then imaged.
Negative control: MCF7 (PMID: 24558488).
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![Western blot - Anti-NNMT antibody [EPR29205-77] - BSA and Azide free (ab318281)](https://www.abcam.cn/ps/products/318/ab318281/Images/ab318281-544360-anti-nnmt-antibody-epr29205-77-western-blot-human-liver-tissue-lysate20-ug.jpg)
Western blot - Anti-NNMT antibody [EPR29205-77] - BSA and Azide free (ab318281)All lanes : Anti-NNMT antibody [EPR29205-77] (ab318280) at 1/1000 dilution
Lane 1 : Human liver tissue lysate
Lane 2 : Human cerebellum tissue lysate
Lane 3 : Human skeletal muscle tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Observed band size: 29 kDa why is the actual band size different from the predicted?
Exposure time: 6 secondsThis data was developed using ab318280, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Negative control: cerebellum, skeletal muscle (PMID: 8182091).
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.
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![Western blot - Anti-NNMT antibody [EPR29205-77] - BSA and Azide free (ab318281)](https://www.abcam.cn/ps/products/318/ab318281/Images/ab318281-544361-anti-nnmt-antibody-epr29205-77-western-blot-huvec-human-umbilical-vein-endothelial-cell-whole-cell-lysate20-ug.jpg)
Western blot - Anti-NNMT antibody [EPR29205-77] - BSA and Azide free (ab318281)All lanes : Anti-NNMT antibody [EPR29205-77] (ab318280) at 1/1000 dilution
Lane 1 : HUVEC (human umbilical vein endothelial cell) whole cell lysate
Lane 2 : A549 (human lung carcinoma epithelial cell) whole cell lysate
Lane 3 : PANC-1 (human pancreatic epithelioid carcinoma epithelial cell) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Observed band size: 29 kDa why is the actual band size different from the predicted?
Exposure time: 6 secondsThis data was developed using ab318280, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Negative control: PANC-1.
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.
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![Western blot - Anti-NNMT antibody [EPR29205-77] - BSA and Azide free (ab318281)](https://www.abcam.cn/ps/products/318/ab318281/Images/ab318281-544362-anti-nnmt-antibody-epr29205-77-western-blot-his-tagged-human-nnmt-recombinant-protein10-ng.jpg)
Western blot - Anti-NNMT antibody [EPR29205-77] - BSA and Azide free (ab318281)All lanes : Anti-NNMT antibody [EPR29205-77] (ab318280) at 1/1000 dilution
Lane 1 : His-tagged human NNMT recombinant protein
Lane 2 : His-tagged human INMT recombinant protein
Lysates/proteins at 0.01 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Observed band size: 29 kDa why is the actual band size different from the predicted?
Exposure time: 10 secondsThis data was developed using ab318280, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
This antibody does not cross-react with human INMT.
In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NNMT antibody [EPR29205-77] - BSA and Azide free (ab318281)](https://www.abcam.cn/ps/products/318/ab318281/Images/ab318281-6-anti-nnmt-antibody-epr29205-77-immunohistochemistry-human-liver.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NNMT antibody [EPR29205-77] - BSA and Azide free (ab318281)This data was developed using ab318280, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling NNMT with ab318280 at 1/2000 (0.253 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Positive staining on human liver. The section was incubated with ab318280 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND™ RX instrument
Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the backgroundCounterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NNMT antibody [EPR29205-77] - BSA and Azide free (ab318281)](https://www.abcam.cn/ps/products/318/ab318281/Images/ab318281-7-anti-nnmt-antibody-epr29205-77-immunohistochemistry-human-colon-carcinom.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NNMT antibody [EPR29205-77] - BSA and Azide free (ab318281)This data was developed using ab318280, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Human colon carcinoma tissue labeling NNMT with ab318280 at 1/2000 (0.253 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Positive staining on stroma of human colon carcinoma. The section was incubated with ab318280 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND™ RX instrument
Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the backgroundCounterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NNMT antibody [EPR29205-77] - BSA and Azide free (ab318281)](https://www.abcam.cn/ps/products/318/ab318281/Images/ab318281-8-anti-nnmt-antibody-epr29205-77-immunohistochemistry-human-breast-carcino.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NNMT antibody [EPR29205-77] - BSA and Azide free (ab318281)This data was developed using ab318280, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labeling NNMT with ab318280 at 1/2000 (0.253 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Positive staining on stroma of human breast carcinoma. The section was incubated with ab318280 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND™ RX instrument
Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the backgroundCounterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NNMT antibody [EPR29205-77] - BSA and Azide free (ab318281)](https://www.abcam.cn/ps/products/318/ab318281/Images/ab318281-10-anti-nnmt-antibody-epr29205-77-immunohistochemistry-awild-type-hela-h.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NNMT antibody [EPR29205-77] - BSA and Azide free (ab318281)This data was developed using ab318280, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded A Wild-type Hela (Human cervix adenocarcinoma) cell pellet B Human NNMT knockout Hela cell pellet tissue labeling NNMT with ab318280 at 1/1000 (0.505 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Positive staining on (A) wild-type Hela cell pellet, no staining on (B) human NNMT knockout Hela cell pellet. The section was incubated with ab318280 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND™ RX instrumentCounterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NNMT antibody [EPR29205-77] - BSA and Azide free (ab318281)](https://www.abcam.cn/ps/products/318/ab318281/Images/ab318281-9-anti-nnmt-antibody-epr29205-77-immunohistochemistry-human-cerebrum.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NNMT antibody [EPR29205-77] - BSA and Azide free (ab318281)This data was developed using ab318280, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling NNMT with ab318280 at 1/2000 (0.253 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Low expression tissue: no staining on human cerebrum. The section was incubated with ab318280 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND™ RX instrument
Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the backgroundCounterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
-
![Immunocytochemistry/ Immunofluorescence - Anti-NNMT antibody [EPR29205-77] - BSA and Azide free (ab318281)](https://www.abcam.cn/ps/products/318/ab318281/Images/ab318281-5-anti-nnmt-antibody-epr29205-77-immunocytochemistry-hela.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-NNMT antibody [EPR29205-77] - BSA and Azide free (ab318281)This data was developed using ab318280, the same antibody clone in a different buffer formulation.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling NNMT with ab318280 at 1/500 (1.01 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).
Confocal image showing nuclear and cytoplasmic staining in parental HeLa cell line (shown in green) and negative staining in NNMT KO HeLa cell line. The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.
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![Anti-NNMT antibody [EPR29205-77] - BSA and Azide free (ab318281)](https://www.abcam.cn/ps/products/318/ab318281/Images/ab318281-11-benefitsofrecombinantantibodies.jpg)
Anti-NNMT antibody [EPR29205-77] - BSA and Azide free (ab318281)
实验方案
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
数据表及文件
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Datasheet download
Certificate of Compliance
文献 (0)
ab318281 尚未被引用在任何文献中。
