Anti-Dnmt1 抗体 (ab19905)
Key features and details
- Rabbit polyclonal to Dnmt1
- Suitable for: WB, ICC/IF, IP, IHC-P
- Knockout validated
- Reacts with: Human
- Isotype: IgG
リコンビナント抗体で、ロット間での高い再現性を実現
- 異なるロット間での安定した再現性
- 容易なスケールアップ
- 評価試験による特異性の確認済み
- 倫理基準に準拠 - アニマル・フリーの生産
製品の概要
-
製品名
Anti-Dnmt1 antibody
Dnmt1 一次抗体 製品一覧 -
製品の詳細
Rabbit polyclonal to Dnmt1 -
由来種
Rabbit -
アプリケーション
適用あり: WB, ICC/IF, IP, IHC-Pmore details -
種交差性
交差種: Human
交差が予測される動物種: Mouse
-
免疫原
Synthetic peptide conjugated to KLH derived from within residues 100 - 200 of Human Dnmt1.
Immunogen の所有権に関して (Peptide available as ab21999.) -
ポジティブ･コントロール
- This antibody gave a positive signal in: (WB) HeLa nuclear; (IF) Hek293; HepG2; MCF7
-
特記事項
The immunogen used to generate this antibody has 71% identity with the corresponding region in mouse Dnmt1. We have had variable feedback about the ability of this antibody to recognise mouse Dnmt1. Customers may prefer to use one of our other antibodies for detection of mouse Dnmt1The 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
-
製品の状態
Liquid -
保存方法
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
バッファー
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help. -
Concentration information loading... -
精製度
Immunogen affinity purified -
ポリ/モノ
ポリクローナル -
アイソタイプ
IgG -
研究分野
関連製品
-
Compatible Secondaries
-
Isotype control
-
Recombinant Protein
-
Related Products
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ab19905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 |
|---|---|---|
| WB | (4) |
Use a concentration of 1 µg/ml. Detects a band of approximately 183 kDa (predicted molecular weight: 183 kDa).
|
| ICC/IF | (1) |
Use a concentration of 1 µg/ml.
|
| IP | (2) |
Use at an assay dependent concentration.
|
| IHC-P | (4) |
Use a concentration of 1 - 10 µg/ml.
|
| 特記事項 |
|---|
|
WB
Use a concentration of 1 µg/ml. Detects a band of approximately 183 kDa (predicted molecular weight: 183 kDa). |
|
ICC/IF
Use a concentration of 1 µg/ml. |
|
IP
Use at an assay dependent concentration. |
|
IHC-P
Use a concentration of 1 - 10 µg/ml. |
ターゲット情報
-
機能
Methylates CpG residues. Preferentially methylates hemimethylated DNA. Associates with DNA replication sites in S phase maintaining the methylation pattern in the newly synthesized strand, that is essential for epigenetic inheritance. Associates with chromatin during G2 and M phases to maintain DNA methylation independently of replication. It is responsible for maintaining methylation patterns established in development. DNA methylation is coordinated with methylation of histones. Mediates transcriptional repression by direct binding to HDAC2. In association with DNMT3B and via the recruitment of CTCFL/BORIS, involved in activation of BAG1 gene expression by modulating dimethylation of promoter histone H3 at H3K4 and H3K9. -
組織特異性
Ubiquitous; highly expressed in fetal tissues, heart, kidney, placenta, peripheral blood mononuclear cells, and expressed at lower levels in spleen, lung, brain, small intestine, colon, liver, and skeletal muscle. Isoform 2 is less expressed than isoform 1. -
配列類似性
Belongs to the C5-methyltransferase family.
Contains 2 BAH domains.
Contains 1 CXXC-type zinc finger. -
ドメイン
The N-terminal part is required for homodimerization and acts as a regulatory domain. -
翻訳後修飾
Sumoylated; sumoylation increases activity. -
細胞内局在
Nucleus. - Information by UniProt
-
参照データベース
- Entrez Gene: 1786 Human
- Entrez Gene: 13433 Mouse
- Omim: 126375 Human
- SwissProt: P26358 Human
- SwissProt: P13864 Mouse
- Unigene: 202672 Human
- Unigene: 128580 Mouse
-
別名
- ADCADN antibody
- AIM antibody
- CXXC finger protein 9 antibody
see all
画像
-
Western blot - Anti-Dnmt1 antibody (ab19905)
Lane 1: Wild type HAP1 whole cell lysate (20 µg)
Lane 2: DNMT1 knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)
Lane 4: Hek293 whole cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab19905 observed at 170 kDa. Red - loading control, ab18058, observed at 130 kDa.ab19905 was shown to recognize DNMT1 when DNMT1 knockout samples were used, along with additional cross-reactive bands. Wild-type and DNMT1 knockout samples were subjected to SDS-PAGE. Ab19905 and ab18058 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 1 ug/ml and 1/10000 dilution respectively. Blots were developed with 800CW Goat anti Rabbit and 680CW Goat anti Mouse secondary antibodies at 1/10010 dilution for 1 hour at room temperature before imaging.
-
Western blot - Anti-Dnmt1 antibody (ab19905)All lanes : Anti-Dnmt1 antibody (ab19905) at 1 µg/ml
Lane 1 : HeLa nuclear lysate
Lane 2 : HeLa nuclear lysate with Human Dnmt1 peptide (ab21999) at 1 µg/ml
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat polyclonal to Rabbit IgG (Alexa Fluor® 680) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 183 kDa
Observed band size: 183 kDa
ab19905 specifically recognises Dnmt1 at 183 kDa in HeLa nuclear extracts (lane1), which is efficiently blocked using the immunising peptide (ab21999) (lane2). -
Immunoprecipitation - Anti-Dnmt1 antibody (ab19905)Dnmt1 was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to Dnmt1 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab19905.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 183kDa: Dnmt1. -
Immunocytochemistry/ Immunofluorescence - Anti-Dnmt1 antibody (ab19905)ab19905 at a 1/200 dilution staining paraformaldehyde fixed asynchronous HeLa cells by ICC/IF. The antibody was incubated with the cells for 30 minutes and then bound antibody was detected using a Cy3 conjugated goat anti-rabbit antibody. Nuclei were visualised using DAPI staining. Ab19905 gives a pattern that is predominantly enriched within nuclei.
This image is courtesy of an Abreview submitted by Kirk McManus.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dnmt1 antibody (ab19905)Staining of human tonsil tissue was performed using ab19905. Strong nuclear staining was observed in germinal center cells and scattered cells in the interfollicular area. T cells were weakly to moderately positive and endothelial cells were positive. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dnmt1 antibody (ab19905)This image is courtesy of an anonymous Abreviewab19905 (1/100) staining Dnmt1 in paraffin-embbeded mouse testis tissue sections. Tissue underwent fixation in formaldehyde, heat-mediated antigen retrieval in 10mM citrate buffer pH 6.0 and blocking (30 minutes, 8% milk). A biotinylated anti-rabbit IgG (H+L) antibody was used as the secondary. For further experimental details please refer to abreview. -
Western blot - Anti-Dnmt1 antibody (ab19905)Jurkat cells were incubated at 37°C for 5 days with vehicle control (0 µM) and different concentrations of hydralazine hydrochloride (ab120863). Decreased expression of DNMT1 (ab19905) in Jurkat cells correlates with an increase in hydralazine hydrochloride concentration, as described in literature.
Whole cell lysates were prepared with RIPA buffer (containing protease inhibitors and sodium orthovanadate), 20µg of each were loaded on the gel and the WB was run under reducing conditions. After transfer the membrane was blocked for an hour using 5% BSA before being incubated with ab19905 at 1 µg/ml and ab8227 at 1 µg/ml overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP (ab97051) at 1/10000 dilution and visualised using ECL development solution.
-
Immunocytochemistry/ Immunofluorescence - Anti-Dnmt1 antibody (ab19905)ICC/IF image of ab19905 stained HeLa cells. The cells were 4% PFA fixed (10 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 (ab19905, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit 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 antibody also gave a positive result in 1% PFA fixed (10 min) Hek293, HepG2 and MCF7 cells at 1µg/ml.
プロトコール
データシートおよび資料
-
Datasheet download
参考文献 (62)
ab19905 は 62 報の論文で使用されています。
- Wang Y et al. Retinoic acid induces NELFA-mediated 2C-like state of mouse embryonic stem cells associates with epigenetic modifications and metabolic processes in chemically defined media. Cell Prolif 54:e13049 (2021). PubMed: 33960560
- Onodera A et al. Roles of TET and TDG in DNA demethylation in proliferating and non-proliferating immune cells. Genome Biol 22:186 (2021). PubMed: 34158086
- Zhao Z et al. LncRNA DLX6-AS1 Promotes Malignant Phenotype and Lymph Node Metastasis in Prostate Cancer by Inducing LARGE Methylation. Front Oncol 10:1172 (2020). PubMed: 32850336
- Cardenas H et al. Methylomic Signatures of High Grade Serous Ovarian Cancer. Epigenetics N/A:1-16 (2020). PubMed: 33289590
- Massah S et al. The effects of the DNA Demethylating reagent, 5-azacytidine on SMCHD1 genomic localization. BMC Genet 21:3 (2020). PubMed: 31941450
