Anti-NMNAT2 抗体 [2E4] (ab56980)
Key features and details
- Mouse monoclonal [2E4] to NMNAT2
- Suitable for: ICC/IF, WB, Sandwich ELISA, IHC-P, Flow Cyt
- Reacts with: Mouse, Human, Recombinant fragment
- Isotype: IgG1
製品の概要
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製品名
Anti-NMNAT2 antibody [2E4]
NMNAT2 一次抗体 製品一覧 -
製品の詳細
Mouse monoclonal [2E4] to NMNAT2 -
由来種
Mouse -
アプリケーション
適用あり: ICC/IF, WB, Sandwich ELISA, IHC-P, Flow Cytmore details -
種交差性
交差種: Mouse, Human, Recombinant fragment -
免疫原
Recombinant fragment corresponding to Human NMNAT2 aa 208-308.
Sequence:CIPGLWNEADMEVIVGDFGIVVVPRDAADTDRIMNHSSILRKYKNNIMVV KDDINHPMSVVSSTKSRLALQHGDGHVVDYLSQPVIDYILKSQLYINASG
Database link: Q9BZQ4 -
ポジティブ・コントロール
- ICC/IF: SHSY5Y cells. IHC-P: Human breast cancer Flow cyt: SH-SY5Y cells.
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特記事項
This product was changed from ascites to tissue culture supernatant on 15th May 2019. 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
製品の特性
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製品の状態
Liquid -
保存方法
Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles. -
バッファー
pH: 7.4
Constituent: PBS -
Concentration information loading...
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精製度
Tissue culture supernatant -
特記事項(精製)
Purified from TCS. -
ポリ/モノ
モノクローナル -
クローン名
2E4 -
アイソタイプ
IgG1 -
軽鎖の種類
kappa -
研究分野
関連製品
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Compatible Secondaries
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Isotype control
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab56980の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
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ICC/IF |
Use at an assay dependent concentration.
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WB | (1) |
Use at an assay dependent concentration. Predicted molecular weight: 34 kDa.
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Sandwich ELISA |
Use at an assay dependent concentration.
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Flow Cyt |
Use at an assay dependent concentration.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
特記事項 |
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ICC/IF
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. Predicted molecular weight: 34 kDa. |
Sandwich ELISA
Use at an assay dependent concentration. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Flow Cyt
Use at an assay dependent concentration. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
ターゲット情報
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機能
Catalyzes the formation of NAD(+) from nicotinamide mononucleotide (NMN) and ATP. Can also use the deamidated form; nicotinic acid mononucleotide (NaMN) as substrate but with a lower efficiency. Cannnot use triazofurin monophosphate (TrMP) as substrate. Also catalyzes the reverse reaction, i.e. the pyrophosphorolytic cleavage of NAD(+). For the pyrophosphorolytic activity prefers NAD(+), NADH and NAAD as substrates and degrades nicotinic acid adenine dinucleotide phosphate (NHD) less effectively. Fails to cleave phosphorylated dinucleotides NADP(+), NADPH and NAADP(+). -
組織特異性
Highly expressed in brain, in particular in cerebrum, cerebellum, occipital lobe, frontal lobe, temporal lobe and putamen. Also found in the heart, skeletal muscle, pancreas and islets of Langerhans. -
パスウェイ
Cofactor biosynthesis; NAD(+) biosynthesis; NAD(+) from nicotinamide D-ribonucleotide: step 1/1. -
配列類似性
Belongs to the eukaryotic NMN adenylyltransferase family. -
細胞内局在
Cytoplasm. Golgi apparatus. - Information by UniProt
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参照データベース
- Entrez Gene: 23057 Human
- Entrez Gene: 226518 Mouse
- Omim: 608701 Human
- SwissProt: Q9BZQ4 Human
- SwissProt: Q8BNJ3 Mouse
- Unigene: 497123 Human
- Unigene: 40548 Mouse
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別名
- C1orf15 antibody
- KIAA0479 antibody
- MGC2756 antibody
see all
画像
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Detection limit for recombinant GST tagged NMNAT2 is 0.03 ng/ml as a capture antibody.
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Western blot against tagged recombinant protein immunogen using ab56980 NMNAT2 antibody at 1ug/ml. Predicted band size of immunogen is 37 kDa
This image was generated using the ascites version of the product.
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ICC/IF image of ab56980 stained SHSY5Y cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab56980, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
This image was generated using the ascites version of the product.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NMNAT2 antibody [2E4] (ab56980)
IHC image of ab56980 staining in human breast cancer formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab56980, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.This image was generated using the ascites version of the product.
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Overlay histogram showing SH-SY5Y cells stained with ab56980 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab56980, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in SH-SY5Y cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.
This image was generated using the ascites version of the product. -
Western blot - Anti-NMNAT2 antibody [2E4] (ab56980)This image is courtesy of an abreview submitted by Jon Gilley, The Babraham Institute, United KingdomAnti-NMNAT2 antibody [2E4] (ab56980) at 2 µg/ml + mouse brain from newborn pup at 40 µg
Secondary
Goat Anti-Mouse IgG (H+L)-HRP Conjugate at 1/3000 dilution
Predicted band size: 34 kDaNmnat2 is a low abundance protein and is difficult to detect. Although multiple non-specific bands are detected by this antibody, we find it is the best commercial antibody for detecting endogenous Nmnat2 as the band is found approximately mid-way between the 25 and 37 kDa markers (at ~32 kDa) and can relatively easily be distinguished from nearby non-specific bands (N.B. a non-specific band at ~37 kDa is also detected in mouse brain extracts from adult mice). Importantly, we do not use this antibody in any immunostaining techniques due to its cross-reactivity with multiple other proteins. The size of the specific band corresponding to endogenous Nmnat2 (32 kDa) was determined based on its migration relative to that of FLAG-tagged Nmnat2 in tranfected HEK cells (which migrates slightly slower at ~34 kDa) and loss of the endogenous Nmnat2 band in transected axons in primary neuronal cultures and in gene trap mouse brain extracts (Gilley and Coleman, PLoS Biology, 2010 [PMID: 20126265]
This image was generated using the ascites version of the product.
プロトコール
データシートおよび資料
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Datasheet download
参考文献 (12)
ab56980 は 12 報の論文で使用されています。
- Tribble JR et al. NAD salvage pathway machinery expression in normal and glaucomatous retina and optic nerve. Acta Neuropathol Commun 11:18 (2023). PubMed: 36681854
- Zhu Y et al. Human Nmnat1 Promotes Autophagic Clearance of Amyloid Plaques in a Drosophila Model of Alzheimer's Disease. Front Aging Neurosci 14:852972 (2022). PubMed: 35401143
- Yagi M et al. Mitochondrial translation deficiency impairs NAD+ -mediated lysosomal acidification. EMBO J 40:e105268 (2021). PubMed: 33528041
- Satoh M et al. A Novel In Vitro Assay Using Human iPSC-Derived Sensory Neurons to Evaluate the Effects of External Chemicals on Neuronal Morphology: Possible Implications in the Prediction of Abnormal Skin Sensation. Int J Mol Sci 22:N/A (2021). PubMed: 34638865
- Liu J et al. NMNAT promotes glioma growth through regulating post-translational modifications of P53 to inhibit apoptosis. Elife 10:N/A (2021). PubMed: 34919052