Anti-5-hydroxymethylcytosine (5-hmC) 抗体 [RM236] (ab214728)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [RM236] to 5-hydroxymethylcytosine (5-hmC)
- Suitable for: ELISA, MeDIP, Flow Cyt, Dot blot, IHC-P, ICC/IF
- Reacts with: Species independent
製品の概要
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製品名
Anti-5-hydroxymethylcytosine (5-hmC) antibody [RM236]
5-hydroxymethylcytosine (5-hmC) 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [RM236] to 5-hydroxymethylcytosine (5-hmC) -
由来種
Rabbit -
特異性
ab214728 reacts to 5-hydroxymethylcytosine in both single-stranded and double-stranded DNA. No cross reactivity with non-methylated cytosine and methylcytosine in DNA. -
アプリケーション
適用あり: ELISA, MeDIP, Flow Cyt, Dot blot, IHC-P, ICC/IFmore details -
種交差性
交差種: Species independent -
免疫原
Chemical/ Small Molecule corresponding to 5-hydroxymethylcytosine (5-hmC) conjugated to Bovine Serum Albumin (BSA).
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ポジティブ・コントロール
- IHC-P: Human brain tissue. ICC/IF: HeLa cells.
製品の特性
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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 long term. Avoid freeze / thaw cycle. -
バッファー
Preservative: 0.09% Sodium azide
Constituents: 50% Glycerol (glycerin, glycerine), 1% BSA, PBS -
Concentration information loading...
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精製度
Protein A purified -
ポリ/モノ
モノクローナル -
クローン名
RM236 -
アイソタイプ
IgG -
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ab214728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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ELISA |
Use a concentration of 0.1 - 1 µg/ml.
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MeDIP |
Use a concentration of 0.2 - 2 µg/ml.
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Flow Cyt |
Use a concentration of 1 - 5 µg/ml.
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Dot blot |
Use a concentration of 0.2 - 1 µg/ml.
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IHC-P |
Use a concentration of 0.1 - 1 µg/ml.
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ICC/IF |
Use a concentration of 0.5 - 2 µg/ml.
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特記事項 |
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ELISA
Use a concentration of 0.1 - 1 µg/ml. |
MeDIP
Use a concentration of 0.2 - 2 µg/ml. |
Flow Cyt
Use a concentration of 1 - 5 µg/ml. |
Dot blot
Use a concentration of 0.2 - 1 µg/ml. |
IHC-P
Use a concentration of 0.1 - 1 µg/ml. |
ICC/IF
Use a concentration of 0.5 - 2 µg/ml. |
ターゲット情報
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関連性
5-Hydroxymethylcytosine (5-hmC) is a modified base form of cytosine recently found in Human/mouse brain and inembryonic stem cells. This DNA pyrimidine nitrogen base can be generated by oxidation of 5-methylcytosine, a reaction mediated by the ten-eleven translocation (TET) family of the 5-mC hydroxylases. The function of this base is still not elucidated but it is believed to play an important role in switching genes on and off. -
別名
- 5-hmC antibody
画像
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Immunohistochemical analysis of paraffin-embedded human brain tissue labeling 5-hydroxymethylcytosine (5-hmC) with ab214728 at 1 μg/mL.
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Immunocytochemical staining of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells using 0.5 μg/mL ab214728 (red). Actin filaments was labeled with fluorescein phalloidin (green). HeLa cells were fixed with 4% paraformaldehyde and permeabilized with methanol (−20 °C) before treatment with 2 N HCl for 30 minutes at 37°C to denature the DNA.
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Direct ELISA of mouse brain genomic DNA using ab214728. The plate was directly coated with different concentrations of genomic DNA isolated from mouse brain tissue. 1 ug/mL or 3 ug/mL of ab214728 was used as the primary antibody and an HRP conjugated anti-rabbit IgG as the secondary antibody.
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hMeDIP was performed using anti-5-hmC antibody (RM236) at a 10:1 DNA:Ab ratio. 1 ng of unmethylated, 5-Methylcytosine (5-mC) or 5-Hydroxymethylcytosine (5-hmC) DNA standard (897 bp) was spiked in 1ug of genomic DNA isolated from HeLa cells as the control. Realtime PCR was then performed to determine the capture of DNA standard as in % of input.
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ELISA of single stranded DNA using ab214728 in a serial dilution. The plate was coated
with streptavidin and then biotinylated single stranded
unmethylated DNA, 5-Methylcytosine (5-mC) DNA, and
5-Hydroxymethylcytosine (5-hmC) DNA. Secondary antibody:
alkaline phosphatase conjugated anti-rabbit IgG
.
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Dot blot of double stranded DNA using ab214728 at 0.2 µg/mL. The membrane was
pre-spotted with 50, 5, and 0.5 ng/dot of double stranded
5-Hydroxymethylcytosine (5-hmC) DNA, 5-Methylcytosine
(5-mC) DNA, and unmethylated DNA. The pre-spotted
membrane was then blotted with ab214728.
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Flow Cytometry analysis of 5-hmC expression in HEK293 cells using ab214728. The cells were fixed with ice-cold MeOH, permeabilized with 0.5% Triton X-100, denatured with 2N HCl, then stained with ab214728 (Blue) or with a negative control antibody (Red).
プロトコール
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (21)
ab214728 は 21 報の論文で使用されています。
- Zachou K et al. Altered DNA methylation pattern characterizes the peripheral immune cells of patients with autoimmune hepatitis. Liver Int 42:1355-1368 (2022). PubMed: 35108441
- Shen Y et al. Loss of 5-hydroxymethylcytosine as a Poor Prognostic Factor for Primary Testicular Diffuse Large B-cell Lymphoma. Int J Med Sci 19:225-232 (2022). PubMed: 35165508
- Kong FC et al. A novel epigenetic marker, Ten-eleven translocation family member 2 (TET2), is identified in the intractable epileptic brain and regulates ATP binding cassette subfamily B member 1 (ABCB1) in the blood-brain barrier. Bioengineered 13:6638-6649 (2022). PubMed: 35235761
- Zhang J et al. Metformin Regulates TET2 Expression to Inhibit Endometrial Carcinoma Proliferation: A New Mechanism. Front Oncol 12:856707 (2022). PubMed: 35480097
- Cao LH et al. Food Allergy-Induced Autism-Like Behavior is Associated with Gut Microbiota and Brain mTOR Signaling. J Asthma Allergy 15:645-664 (2022). PubMed: 35603013