重组Anti-SIRT6抗体[EPR26255-85] (ab289970)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR26255-85] to SIRT6
- Suitable for: Flow Cyt (Intra), IHC-P, WB, ICC/IF, IP
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
概述
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产品名称
Anti-SIRT6抗体[EPR26255-85]
参阅全部 SIRT6 一抗 -
描述
兔单克隆抗体[EPR26255-85] to SIRT6 -
宿主
Rabbit -
经测试应用
适用于: Flow Cyt (Intra), IHC-P, WB, ICC/IF, IPmore details -
种属反应性
与反应: Human -
免疫原
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: HeLa, HepG2, MCF7, HCT116 and Jurkat whole cell lysates; Human kidney tissue lysate. IHC-P: Human colon, thymus tissue; HeLa cell pellet.. Flow Cyt (intra): HeLa cells. IP: HeLa whole cell lysate. ICC/IF : Wild-type 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: 7.20
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
EPR26255-85 -
同种型
IgG -
研究领域
相关产品
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Compatible Secondaries
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Isotype control
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab289970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) |
1/5000.
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IHC-P |
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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WB |
1/1000. Predicted molecular weight: 39 kDa.
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ICC/IF |
1/1000.
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IP |
1/30.
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说明 |
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Flow Cyt (Intra)
1/5000. |
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
WB
1/1000. Predicted molecular weight: 39 kDa. |
ICC/IF
1/1000. |
IP
1/30. |
靶标
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功能
NAD-dependent protein deacetylase. Has deacetylase activity towards histone H3K9Ac and H3K56Ac. Modulates acetylation of histone H3 in telomeric chromatin during the S-phase of the cell cycle. Deacetylates histone H3K9Ac at NF-kappa-B target promoters and may down-regulate the expression of a subset of NF-kappa-B target genes. Acts as a corepressor of the transcription factor HIF1A to control the expression of multiple glycolytic genes to regulate glucose homeostasis. Required for genomic stability. Regulates the production of TNF protein. Has a role in the regulation of life span (By similarity). Deacetylation of nucleosomes interferes with RELA binding to target DNA. May be required for the association of WRN with telomeres during S-phase and for normal telomere maintenance. Required for genomic stability. Required for normal IGF1 serum levels and normal glucose homeostasis. Modulates cellular senescence and apoptosis. On DNA damage, promotes DNA end resection via deacetylation of RBBP8. Has very weak deacetylase activity and can bind NAD(+) in the absence of acetylated substrate. -
序列相似性
Belongs to the sirtuin family. Class IV subfamily.
Contains 1 deacetylase sirtuin-type domain. -
细胞定位
Nucleus, nucleoplasm. Predominantly nuclear. Associated with telomeric heterochromatin regions. - Information by UniProt
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数据库链接
- Entrez Gene: 51548 Human
- Omim: 606211 Human
- SwissProt: Q8N6T7 Human
- Unigene: 423756 Human
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别名
- 2810449N18Rik antibody
- AI043036 antibody
- Mono ADP ribosyltransferase sirtuin 6 antibody
see all
图片
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All lanes : Anti-SIRT6 antibody [EPR26255-85] (ab289970) at 1/1000 dilution
Lane 1 : Wild-type HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : SIRT6 knockout HeLa whole cell lysate
Lane 3 : Jurkat (human T cell leukemia T lymphocyte) whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 39 kDa
Observed band size: 40, 43 kDa why is the actual band size different from the predicted?Blocking buffer and concentration: Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS.
Diluting buffer and concentration: Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBST.Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
False colour image of Western blot: Anti-SIRT6 antibody [EPR26255-85](ab289970) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red.
In Western blot, ab289970 was shown to bind specifically to SIRT6. Two bands were observed at 43/36 kDa in wild-type SIRT6 cell lysates with no signal observed at this size in SIRT6 knockout cell line ab265054 (knockout cell lysate ab257673). To generate this image, wild-type and SIRT6 knockout HeLa cell lysates were analyzed. First, samples were run on an SDS-PAGE gel then transferred onto an immobilon-FL PVDF membrane. Membranes were blocked in Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/10000 dilution.
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Lane 1 : Anti-SIRT6 antibody [EPR26255-85] (ab289970)
Lanes 2-4 : Anti-SIRT6 antibody [EPR26255-85] (ab289970) at 1/1000 dilution
Lane 1 : Human kidney tissue lysate
Lane 2 : HepG2 (human hepatocellar carcinoma epithelial cell) whole cell lysate
Lane 3 : MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate
Lane 4 : HCT116 (human colorectal carcinoma epithelial cell) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Predicted band size: 39 kDa
Observed band size: 40, 43 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST
Exposure times: Lane 1: 26 seconds Lane 2-4: 7.75 seconds
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRT6 antibody [EPR26255-85] (ab289970)
Immunohistochemical analysis of paraffin-embedded Human colon tissue labelling SIRT6 with ab289970 at 1/2000 followed by LeicaDS9800 (Bond™, Polymer Refine Detection). Positive staining on human colon (PMID 30333863) (PMID 29387864). The section was incubated with ab289970 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 LeicaDS9800 (Bond™, Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRT6 antibody [EPR26255-85] (ab289970)
Immunohistochemical analysis of paraffin-embedded Parental HeLa cell pellet labelling SIRT6 with ab289970 at 1/10000 followed by LeicaDS9800 (Bond™, Polymer Refine Detection) was used. Positive staining on Parental HeLa cell pellet (A), no staining on SIRT6 KO HeLa cell pellet (B). The section was incubated with ab289970 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 LeicaDS9800 (Bond™, Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRT6 antibody [EPR26255-85] (ab289970)
Immunohistochemical analysis of paraffin-embedded Human thymus tissue labelling SIRT6 with ab289970 at 1/2000 followed by LeicaDS9800 (Bond™, Polymer Refine Detection). Positive staining on human thymus (PMID 29387864). The section was incubated with ab289970 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 LeicaDS9800 (Bond™, Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
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ab289970 staining SIRT6 in wild-type HeLa (human cervix adenocarcinoma epithelial cell line) cells and SIRT6 knockout HeLa (human cervix adenocarcinoma epithelial cell line) cells (ab265054). The cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. The cells were then incubated with ab289970 at 1/1000 dilution and ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) at 1/200 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) secondary antibody at 1/1000 dilution. Nuclear DNA was labelled in blue with DAPI. Confocal image showing nuclear staining in parental Hela cell line and no staining in SIRT6 KO Hela cell line.
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Flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized Parental HeLa (Human cervix adenocarcinoma epithelial cell, Right) / SIRT6 KO HeLa (Left) cells labelling SIRT6 with ab289970 at 1/5000 dilution (0.01µg) (Red) 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, ab150081) at 1/2000 dilution was used as the secondary antibody. Positive staining on HeLa cells (ab255448), while no staining on SIRT6 knockout HeLa cells (ab265054).
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SIRT6 was immunoprecipitated from HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate with ab289970 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab289970 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10 µg
Lane 2: ab289970 IP in HeLa whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab289970 in HeLa whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 7.75 seconds
实验方案
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.
数据表及文件
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SDS download
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Datasheet download
Certificate of Compliance
文献 (1)
ab289970 被引用在 1 文献中.
- Xie T et al. Luteolin suppresses TNF-a-induced inflammatory injury and senescence of nucleus pulposus cells via the Sirt6/NF-?B pathway. Exp Ther Med 24:469 (2022). PubMed: 35747154