Anti-KDM3A / JHDM2A 抗体 [EPR18875-106] (ab243641)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR18875-106] to KDM3A / JHDM2A
- Suitable for: Flow Cyt (Intra), IHC-P, IP, WB, ICC/IF
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
製品の概要
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製品名
Anti-KDM3A / JHDM2A antibody [EPR18875-106]
KDM3A / JHDM2A 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EPR18875-106] to KDM3A / JHDM2A -
由来種
Rabbit -
アプリケーション
適用あり: Flow Cyt (Intra), IHC-P, IP, WB, ICC/IFmore details
適用なし: ChIP -
種交差性
交差種: Mouse, Rat, Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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ポジティブ・コントロール
- WB: HeLa, HeLa, Ramos, NIH/3T3 and C6 whole cell lysates. IHC-P: Human testis tissue; Mouse testis tissue; Rat testis tissue. ICC/IF: HeLa cells. Flow Cyt (intra): HeLa cells treated with 1mM DFO for 24h.IP: Ramos whole cell lysate.
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特記事項
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.
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
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
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
EPR18875-106 -
アイソタイプ
IgG -
研究分野
関連製品
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Alternative Versions
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Compatible Secondaries
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Isotype control
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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ab243641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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Flow Cyt (Intra) |
1/500.
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IHC-P |
1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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IP |
1/30.
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WB |
1/1000. Detects a band of approximately 60, 90, 147 kDa (predicted molecular weight: 147 kDa).
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ICC/IF |
1/100.
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特記事項 |
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Flow Cyt (Intra)
1/500. |
IHC-P
1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
IP
1/30. |
WB
1/1000. Detects a band of approximately 60, 90, 147 kDa (predicted molecular weight: 147 kDa). |
ICC/IF
1/100. |
ターゲット情報
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機能
Histone demethylase that specifically demethylates 'Lys-9' of histone H3, thereby playing a central role in histone code. Preferentially demethylates mono- and dimethylated H3 'Lys-9' residue, with a preference for dimethylated residue, while it has weak or no activity on trimethylated H3 'Lys-9'. Demethylation of Lys residue generates formaldehyde and succinate. Involved in hormone-dependent transcriptional activation, by participating in recruitment to androgen-receptor target genes, resulting in H3 'Lys-9' demethylation and transcriptional activation. Involved in spermatogenesis by regulating expression of target genes such as PRM1 and TMP1 which are required for packaging and condensation of sperm chromatin. Involved in obesity resistance through regulation of metabolic genes such as PPARA and UCP1. -
配列類似性
Belongs to the JHDM2 histone demethylase family.
Contains 1 JmjC domain. -
ドメイン
The JmjC domain and the C6-type zinc-finger are required for the demethylation activity.
Leu-Xaa-Xaa-Leu-Leu (LXXLL) motifs are known to mediate the association with nuclear receptors. -
細胞内局在
Cytoplasm. Nucleus. Nuclear in round spermatids. When spermatids start to elongate, localizes to the cytoplasm where it forms distinct foci which disappear in mature spermatozoa. - Information by UniProt
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参照データベース
- Entrez Gene: 55818 Human
- Entrez Gene: 104263 Mouse
- Entrez Gene: 312440 Rat
- Omim: 611512 Human
- SwissProt: Q9Y4C1 Human
- SwissProt: Q6PCM1 Mouse
- SwissProt: Q63679 Rat
- Unigene: 557425 Human
see all -
別名
- DKFZp686A24246 antibody
- DKFZp686P07111 antibody
- JHDM2A antibody
see all
画像
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Immunohistochemical analysis of paraffin-embedded Rat testis tissue labeling KDM3A / JHDM2A with ab243641 at 1/500 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on rat testis (PMID: 17943087). The section was incubated with ab243641 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.
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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa cells labelling KDM3A / JHDM2A with ab243641 at 1/100 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing nuclear staining in HeLa cell line treatment with DFO (1mM) for 24h. ab195889 Anti-alpha Tubulin antibody (Alexa Fluor® 594) was used to counterstain tubulin at 1/1000 dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.
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Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized HeLa cells treated with 1mM DFO for 24h (Red) / Untreated control (Green) labelling KDM3A / JHDM2A with ab243641 at 1/500 dilution (0.1ug) (Red) / (Green) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
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KDM3A / JHDM2A was immunoprecipitated from 0.35 mg Ramos (human Burkitt's lymphoma B lymphocyte), whole cell lysate with ab243641 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab243641 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: Ramos (human Burkitt's lymphoma B lymphocyte), whole cell lysate 10 ug
Lane 2: ab243641 IP in Ramos whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab243641 in Ramos whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 minutes.
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All lanes : Anti-KDM3A / JHDM2A antibody [EPR18875-106] (ab243641) at 1/1000 dilution
Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 20 µg
Lane 2 : HeLa treated with 1mM Deferoxamine mesylate (DFO) for 24 hours, whole cell lysate
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution
Predicted band size: 147 kDa
Observed band size: 147 kDaBlocking and diluting buffer and concentration: 5% NFDM/TBST.
Fresh lysates were used in this WB.
Exposure time: 3 minutes.
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Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labeling KDM3A / JHDM2A with ab243641 at 1/500 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse testis (PMID: 17943087). The section was incubated with ab243641 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.
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Anti-KDM3A / JHDM2A antibody [EPR18875-106] (ab243641) at 1/1000 dilution + NIH/3T3 (mouse embryonic fibroblast), whole cell lysate at 20 µg
Secondary
Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution
Predicted band size: 147 kDa
Observed band size: 147,60,90 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST.
This blot was developed using a higher sensitivity ECL substrate.
Fresh lysates were used in this WB.
The bands at 90, 60KD are the degragated fragments of KDM3A which is consistent with the literature (PMID: 31629659, 26728187).
Exposure time: 3 minutes.
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Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling KDM3A / JHDM2A with ab243641 at 1/500 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on human testis (PMID: 25071150). The section was incubated with ab243641 for 30 mins at room temperature.T he immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.
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Anti-KDM3A / JHDM2A antibody [EPR18875-106] (ab243641) at 1/1000 dilution + Ramos (human Burkitts lymphoma B lymphocyte), whole cell lysate at 30 µg
Secondary
VeriBlot for IP secondary antibody(HRP)(ab131366) at 1/5000 dilution
Predicted band size: 147 kDa
Observed band size: 147 kDaBlocking and diluting buffer and concentration: 5% NFDM/TBST.
This blot was developed using a higher sensitivity ECL substrate.
Exposure time: 3 minutes.
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Anti-KDM3A / JHDM2A antibody [EPR18875-106] (ab243641) at 1/1000 dilution + C6 (rat glial tumor glial cell), whole cell lysate at 20 µg
Secondary
Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution
Predicted band size: 147 kDa
Observed band size: 147,60 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST.
This blot was developed using a higher sensitivity ECL substrate.
Fresh lysates were used in this WB.
The band at 60KD is the degragated fragment of KDM3A which is consistent with the literature(PMID: 31629659, 26728187).
Exposure time: 3 minutes.
プロトコール
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.
データシートおよび資料
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SDS download
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Datasheet download
Certificate of Compliance
参考文献 (1)
ab243641 は 1 報の論文で使用されています。
- Liu J et al. Histone Demethylase KDM3A Promotes Cervical Cancer Malignancy Through the ETS1/KIF14/Hedgehog Axis. Onco Targets Ther 13:11957-11973 (2020). PubMed: 33239895