重组Anti-mH2A1抗体[EPR9359(2)] (ab183041)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR9359(2)] to mH2A1
- Suitable for: WB, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Mouse, Human
Related conjugates and formulations
概述
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产品名称
Anti-mH2A1抗体[EPR9359(2)]
参阅全部 mH2A1 一抗 -
描述
兔单克隆抗体[EPR9359(2)] to mH2A1 -
宿主
Rabbit -
经测试应用
适用于: WB, IHC-P, ICC/IFmore details -
种属反应性
与反应: Mouse, Human
预测可用于: Rat -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: HepG2, MCF7, 293T and HeLa whole cell lysate (ab150035) IHC-P: Human kidney and liver tissues ICC-IF: HAP1-WT and H2AFY knockout cells. MCF7 and HeLa cells.
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常规说明
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 short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
存储溶液
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR9359(2) -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
- Alexa Fluor® 488 Anti-mH2A1 antibody [EPR9359(2)] (ab208563)
- Alexa Fluor® 647 Anti-mH2A1 antibody [EPR9359(2)] (ab208879)
- HRP Anti-mH2A1 antibody [EPR9359(2)] (ab209320)
- Alexa Fluor® 555 Anti-mH2A1 antibody [EPR9359(2)] (ab211851)
- Alexa Fluor® 594 Anti-mH2A1 antibody [EPR9359(2)] (ab217090)
- Anti-mH2A1 antibody [EPR9359(2)] - BSA and Azide free (ab232602)
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
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Recombinant Protein
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab183041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 |
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WB |
1/10000 - 1/50000. Detects a band of approximately 40 kDa (predicted molecular weight: 40 kDa).
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IHC-P | (1) |
1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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ICC/IF | (1) |
Use a concentration of 1 µg/ml.
This antibody gives positive signal in both 4%PFA and 100% MeOH-fixed cells. |
说明 |
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WB
1/10000 - 1/50000. Detects a band of approximately 40 kDa (predicted molecular weight: 40 kDa). |
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
ICC/IF
Use a concentration of 1 µg/ml. This antibody gives positive signal in both 4%PFA and 100% MeOH-fixed cells. |
靶标
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功能
Variant histone H2A which replaces conventional H2A in a subset of nucleosomes where it represses transcription. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling. Involved in stable X chromosome inactivation. Inhibits the binding of transcription factors and interferes with the activity of remodeling SWI/SNF complexes. Inhibits histone acetylation by EP300 and recruits class I HDACs, which induces an hypoacetylated state of chromatin. In addition, isoform 1, but not isoform 2, binds ADP-ribose and O-acetyl-ADP-ribose, and may be involved in ADP-ribose-mediated chromatin modulation. -
组织特异性
Ubiquitous. -
序列相似性
Contains 1 histone H2A domain.
Contains 1 Macro domain. -
翻译后修饰
Monoubiquitinated at either Lys-116 or Lys-117. May also be polyubiquitinated. Ubiquitination is mediated by the CUL3/SPOP E3 complex and does not promote proteasomal degradation. Instead, it is required for enrichment in inactive X chromosome chromatin. -
细胞定位
Nucleus. Chromosome. Enriched in inactive X chromosome chromatin and in senescence-associated heterochromatin. - Information by UniProt
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数据库链接
- Entrez Gene: 9555 Human
- Entrez Gene: 26914 Mouse
- Entrez Gene: 29384 Rat
- Omim: 610054 Human
- SwissProt: O75367 Human
- SwissProt: Q9QZQ8 Mouse
- SwissProt: Q02874 Rat
- Unigene: 420272 Human
see all -
别名
- Core histone macro h2a.1 antibody
- Core histone macro-H2A.1 antibody
- H2A histone family member Y antibody
see all
图片
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Lane 1: Wild type HAP1 whole cell lysate (20 µg)
Lane 2: mH2A1 knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)
Lane 4: Hepg2 whole cell lysate (20 µg)Lanes 1 - 4: Merged signal (red and green). Green - ab183041 observed at 40 kDa. Red - loading control, ab18058, observed at 130 kDa.
ab183041 was shown to specifically react with mH2A1 when mH2A1 knockout samples were used. Wild-type and mH2A1 knockout samples were subjected to SDS-PAGE. Ab183041 and ab18058 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 10000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
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ab183041 staining mH2A1 in HAP1 WT and H2AFY knockout cells. The cells were fixed with 100% methanol (5min), permeabilized with 0.1%PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab183041 at 1μg/ml and ab195889, Mouse monoclonal [DM1A] to alpha Tubulin - Microtubule Marker (Alexa Fluor® 594) at 1/250 dilution (shown in pseudocolor red). Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) at 1/1000 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
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Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling mH2A1 with ab183041 at 1/100 dilution followed by prediluted HRP Polymer for Rabbit IgG. Counter stained with Hematoxylin.
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All lanes : Anti-mH2A1 antibody [EPR9359(2)] (ab183041) at 1/1000 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : H2AFY CRISPR/Cas9 edited HEK-293T cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 40 kDa
Observed band size: 40 kDaLanes 1- 2: Merged signal (red and green). Green - ab183041 observed at 40 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.
ab183041 was shown to react with mH2A1 in wild-type HEK-293T cells in western blot. The band observed in CRISPR/Cas9 edited cell line ab266241 (CRISPR/Cas9 edited cell lysate ab257463) lane below 40kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HEK-293T and H2AFY CRISPR/Cas9 edited HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab183041 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at a 1 in 1000 Dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-mH2A1 antibody [EPR9359(2)] (ab183041) at 1/50000 dilution
Lane 1 : HepG2 cell lysate
Lane 2 : MCF7 cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : 293T cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugate at 1/1000 dilution
Predicted band size: 40 kDa
Observed band size: 40 kDa -
Immunofluorescent analysis of 4% paraformaldehyde-fixed MCF7 cells labeling mH2A1 with ab183041 at 1/250 dilution followed by Goat anti rabbit IgG (Alexa Fluor® 488) at 1/200 dilution (green). Counter stained with Dapi (blue).
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Immunofluorescent analysis of acetone-fixed HeLa cells labeling mH2A1 with ab183041 at 1/250 dilution followed by Goat anti rabbit IgG (Alexa Fluor® 488) at 1/200 dilution (green). Counter stained with Dapi (blue).
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Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling mH2A1 with ab183041 at 1/100 dilution followed by prediluted HRP Polymer for Rabbit IgG. Counter stained with Hematoxylin.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse liver from wild-type and mH2A1/2 knock out tissue sections labeling mH2A1 with ab183041 at 1/400 dilution. Sections were fixed in formaldehyde; heat mediated antigen retivial was performed using a citrate buffer pH 6. An undiluted polyclonal horse anti-rabbit IgG (HRP-conjugated) was used as the secondary antibody.
实验方案
数据表及文件
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SDS download
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Datasheet download
Certificate of Compliance
文献 (13)
ab183041 被引用在 13 文献中.
- Mattola S et al. Parvovirus nonstructural protein 2 interacts with chromatin-regulating cellular proteins. PLoS Pathog 18:e1010353 (2022). PubMed: 35395063
- Xu X et al. The epigenetic regulator LSH maintains fork protection and genomic stability via MacroH2A deposition and RAD51 filament formation. Nat Commun 12:3520 (2021). PubMed: 34112784
- Wang Y et al. Genome oligopaint via local denaturation fluorescence in situ hybridization. Mol Cell 81:1566-1577.e8 (2021). PubMed: 33657402
- Ni K & Muegge K LSH catalyzes ATP-driven exchange of histone variants macroH2A1 and macroH2A2. Nucleic Acids Res 49:8024-8036 (2021). PubMed: 34223906
- Cheng Y et al. TAD-like single-cell domain structures exist on both active and inactive X chromosomes and persist under epigenetic perturbations. Genome Biol 22:309 (2021). PubMed: 34749781