重组Anti-Histone H2A.Z抗体[EPR6171(2)(B)] - ChIP Grade (ab150402)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR6171(2)(B)] to Histone H2A.Z - ChIP Grade
- Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, ChIP
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-Histone H2A.Z抗体[EPR6171(2)(B)] - ChIP Grade
参阅全部 Histone H2A.Z 一抗 -
描述
兔单克隆抗体[EPR6171(2)(B)] to Histone H2A.Z - ChIP Grade -
宿主
Rabbit -
经测试应用
适用于: Flow Cyt (Intra), WB, IHC-P, ICC/IF, ChIPmore details -
种属反应性
与反应: Mouse, Rat, Human -
免疫原
Synthetic peptide within Human Histone H2A.Z aa 1-100. The exact sequence is proprietary.
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阳性对照
- WB: Neuro-2a, HeLa, HepG2, RAW 264.7, C6 and PC-12 cell lysates. IHC-P: Human colon and lung carcinoma tissues, human breast tissue, mouse and rat cerebrum tissue. ICC/IF: HepG2 and HeLa cells. Flow Cyt (intra): 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
Preservative: 0.01% Sodium azide
Constituents: 59% 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
EPR6171(2)(B) -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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ChIP Related Products
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Isotype control
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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ab150402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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Flow Cyt (Intra) |
1/2500.
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WB | (5) |
1/1000 - 1/10000. Predicted molecular weight: 13 kDa.
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IHC-P |
1/2000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
See IHC antigen retrieval protocols. For unpurified format use at 1/250 - 1/500 dilution. |
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ICC/IF |
1/100 - 1/250.
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ChIP |
Use 2 µg for 25 µg of chromatin.
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说明 |
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Flow Cyt (Intra)
1/2500. |
WB
1/1000 - 1/10000. Predicted molecular weight: 13 kDa. |
IHC-P
1/2000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. See IHC antigen retrieval protocols. For unpurified format use at 1/250 - 1/500 dilution. |
ICC/IF
1/100 - 1/250. |
ChIP
Use 2 µg for 25 µg of chromatin. |
靶标
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功能
Variant histone H2A which replaces conventional H2A in a subset of nucleosomes. 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. May be involved in the formation of constitutive heterochromatin. May be required for chromosome segregation during cell division. -
序列相似性
Belongs to the histone H2A family. -
翻译后修饰
Monoubiquitination of Lys-122 gives a specific tag for epigenetic transcriptional repression.
Acetylated on Lys-5, Lys-8 and Lys-12 during interphase. Acetylation disappears at mitosis.
Monomethylated on Lys-5 and Lys-8 by SETD6. SETD6 predominantly methylates Lys-8, lys-5 being a possible secondary site.
Not phosphorylated. -
细胞定位
Nucleus. Chromosome. - Information by UniProt
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数据库链接
- Entrez Gene: 3015 Human
- Entrez Gene: 51788 Mouse
- Entrez Gene: 58940 Rat
- Omim: 142763 Human
- SwissProt: P0C0S5 Human
- SwissProt: P0C0S6 Mouse
- SwissProt: P0C0S7 Rat
- Unigene: 119192 Human
see all -
别名
- H2A histone family member Z antibody
- H2A.z antibody
- H2A/z antibody
see all
图片
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All lanes : Anti-Histone H2A.Z antibody [EPR6171(2)(B)] - ChIP Grade (ab150402) at 1/5000 dilution (Purified)
Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
Lane 2 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysates
Lane 3 : Neuro-2a (Mouse neuroblastoma neuroblast) whole cell lysates
Lane 4 : RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysates
Lane 5 : C6 (Rat glial tumor glial cell) whole cell lysates
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 13 kDa
Observed band size: 13 kDa -
All lanes : Anti-Histone H2A.Z antibody [EPR6171(2)(B)] - ChIP Grade (ab150402) at 1/1000 dilution
Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates prepared in RIPA lysis method
Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates prepared in 1% SDS Hot lysis method
Lane 3 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysates prepared in RIPA lysis method
Lane 4 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysates prepared in 1% SDS Hot lysis method
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 13 kDa
Observed band size: 13 kDaBlocking buffer: 5% NFDM/TBST.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human breast tissue sections labeling Histone H2A.Z with purified ab150402 at 1/2000 dilution (1.09 µg/ml). Heat mediated antigen retrieval was performed using heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
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Chromatin was prepared from HeLa cells according to the Abcam X-ChIP protocol*. Cells were fixed with formaldehyde for 10 minutes.
The ChIP was performed with 25 µg of chromatin, 2 µg of ab150402 (red), and 20 µl of Protein A/G sepharose beads. 2 µg of rabbit normal IgG was added to the beads control (gray). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci, Sybr green approach for heterochromatic loci).
Primers and probes are located in the first kb of the transcribed region.
*http://www.abcam.com/resources?keywords=X%20ChIP%20protocol -
Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Histone H2A.Z with purified ab150402 at 1/200 dilution (10 µg/ml). Cells were fixed in 100% Methanol. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
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Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling Histone H2A.Z (red) with purified ab150402 at a 1/2500 dilution. Cells were fixed with 80% methanol and permeabilized with 0.1% Tween-20. A goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (ab172730). Blue (unlabeled control) - Cells without incubation with the primary and secondary antibodies.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Rat cerebrum tissue sections labeling Histone H2A.Z with purified ab150402 at 1:2000 dilution (1.09 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse cerebrum tissue sections labeling Histone H2A.Z with purified ab150402 at 1/2000 dilution (1.09 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
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Chromatin was prepared from HeLa (Human epithelial cell line from cervix adenocarcinoma) cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 2µg of ab150402 (unpurified) (blue), and 20µl of Anti rabbit IgG sepharose beads. 2μg of rabbit normal IgG was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach). Primers and probes are located in the first kb of the transcribed region.
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Immunohistochemical analysis of paraffin-embedded human colon tissue labeling Histone H2A.Z with ab150402 (unpurified) at 1/250 dilution.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Immunofluorescent analysis of HepG2 (Human liver hepatocellular carcinoma cell line) cells labeling Histone H2A.Z with ab150402 (unpurified) at 1/100 dilution.
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All lanes : Anti-Histone H2A.Z antibody [EPR6171(2)(B)] - ChIP Grade (ab150402) at 1/1000 dilution (unpurified)
Lane 1 : Recombinant Human octamers containing H2A at 1 µg
Lane 2 : Recombinant Human octamers containing H2A at 0.5 µg
Lane 3 : Native recombinant octamers K562 cells at 3 µg
Lane 4 : Native recombinant octamers K562 cells at 1.5 µg
Lane 5 : Recombinant Human octamers containing H2A.Z.2.1 at 0.5 µg
Lane 6 : Recombinant Human octamers containing H2A.Z.1 at 0.5 µg
Secondary
All lanes : HRP-conjugated donkey anti-rabbit IgG polyclonal at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 13 kDa
Observed band size: 15 kDa why is the actual band size different from the predicted?
Exposure time: 5 minutes
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Immunohistochemical analysis of paraffin-embedded human lung carcinoma tissue labeling Histone H2A.Z with ab150402 (unpurified) at 1/250 dilution.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin embedded human cervical carcinoma tissue using ab150402 (unpurified) showing positive staining.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin embedded normal human kidney tissue using ab150402 (unpurified) showing positive staining.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin embedded human prostate hyperplasia tissue using ab150402 showing positive staining.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin embedded human ovarian carcinoma tissue using ab150402 (unpurified) showing positive staining.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
实验方案
数据表及文件
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SDS download
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Datasheet download
文献 (8)
ab150402 被引用在 8 文献中.
- Li Z et al. H2A Histone Family Member Z (H2AFZ) Serves as a Prognostic Biomarker in Lung Adenocarcinoma: Bioinformatic Analysis and Experimental Validation. Med Sci Monit 28:e933447 (2022). PubMed: 35027526
- Wimmers F et al. The single-cell epigenomic and transcriptional landscape of immunity to influenza vaccination. Cell 184:3915-3935.e21 (2021). PubMed: 34174187
- Thomsen I et al. RUNX1 Regulates a Transcription Program That Affects the Dynamics of Cell Cycle Entry of Naive Resting B Cells. J Immunol 207:2976-2991 (2021). PubMed: 34810221
- Lowe DJ et al. Chronic irradiation of human cells reduces histone levels and deregulates gene expression. Sci Rep 10:2200 (2020). PubMed: 32042076
- Yang B et al. H2A.Z regulates tumorigenesis, metastasis and sensitivity to cisplatin in intrahepatic cholangiocarcinoma. Int J Oncol 52:1235-1245 (2018). PubMed: 29532867
- Cheung P et al. Single-Cell Chromatin Modification Profiling Reveals Increased Epigenetic Variations with Aging. Cell 173:1385-1397.e14 (2018). PubMed: 29706550
- Nakamura K et al. Running training experience attenuates disuse atrophy in fast-twitch skeletal muscles of rats. J Appl Physiol (1985) 123:902-913 (2017). PubMed: 28775067
- Weyemi U et al. The histone variant H2A.X is a regulator of the epithelial-mesenchymal transition. Nat Commun 7:10711 (2016). WB ; Human . PubMed: 26876487