使用敲除细胞株进行验证重组RabMAb

重组Anti-SIRT6抗体[EPR18463] (ab191385)

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Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR18463] to SIRT6
  • Suitable for: ICC/IF, IP, WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

概述

  • 产品名称

    Anti-SIRT6抗体[EPR18463]
    参阅全部 SIRT6 一抗

  • 描述

    兔单克隆抗体[EPR18463] to SIRT6

  • 宿主

    Rabbit

  • 经测试应用

    适用于: ICC/IF, IP, WBmore details

  • 种属反应性

    与反应: Mouse, Rat, Human

  • 免疫原

    Synthetic peptide within Human SIRT6 aa 1-100. The exact sequence is proprietary.
    Database link: Q8N6T7

  • 阳性对照

    • WB: HeLa, Jurkat, NIH/3T3, C6, RAW 264.7 and PC-12 cell lysates; HeLa nuclear lysate; rat brain and spleen lysates. ICC/IF: HeLa and HCT 116 cells. IP: Jurkat whole cell lysate.

  • 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
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

性能

应用

Our Abpromise guarantee covers the use of ab191385 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
ICC/IF 1/1000.
IP 1/40.
WB 1/2000. Detects a band of approximately 42 kDa (predicted molecular weight: 39 kDa).

靶标

  • 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

  • 数据库链接

    see all

  • 别名

    • 2810449N18Rik antibody
    • AI043036 antibody
    • Mono ADP ribosyltransferase sirtuin 6 antibody
    • NAD-dependent protein deacetylase sirtuin-6 antibody
    • Regulatory protein SIR2 homolog 6 antibody
    • Regulatory protein SIR2 homolog antibody
    • SIR2 like 6 antibody
    • SIR2 like protein 6 antibody
    • Sir2 related protein type 6 antibody
    • SIR2-like protein 6 antibody
    • SIR2L6 antibody
    • SIR6_HUMAN antibody
    • SIRT 6 antibody
    • SIRT6 antibody
    • Sirtuin (silent mating type information regulation 2 homolog) 6 (S. cerevisiae) antibody
    • Sirtuin 6 antibody
    • Sirtuin type 6 antibody
    • Sirtuin6 antibody
    see all

图片

  • Western blot - Anti-SIRT6 antibody [EPR18463] (ab191385)
    Western blot - Anti-SIRT6 antibody [EPR18463] (ab191385)
    All lanes : Anti-SIRT6 antibody [EPR18463] (ab191385) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : SIRT6 knockout HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 39 kDa
    Observed band size: 42 kDa
    why is the actual band size different from the predicted?



    Lanes 1-2: Merged signal (red and green). Green - ab191385 observed at 40 kDa. Red - loading control ab7291 observed at 50 kDa.

     ab191385 Anti-SIRT6 antibody [EPR18463] was shown to specifically react with SIRT6 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265054 (knockout cell lysate ab257673) was used. Wild-type and SIRT6 knockout samples were subjected to SDS-PAGE. ab191385 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 1000 Dilution and 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.

     

     

  • Western blot - Anti-SIRT6 antibody [EPR18463] (ab191385)
    Western blot - Anti-SIRT6 antibody [EPR18463] (ab191385)
    Anti-SIRT6 antibody [EPR18463] (ab191385) at 1/10000 dilution + HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate at 20 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

    Predicted band size: 39 kDa
    Observed band size: 42 kDa why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The observed MW and doublets are consistent with what has been described in the literature. Two bands run closely together as  doublets representing distinct isoforms; see UniProt annotation and PMID 24169447.

  • Western blot - Anti-SIRT6 antibody [EPR18463] (ab191385)
    Western blot - Anti-SIRT6 antibody [EPR18463] (ab191385)
    Anti-SIRT6 antibody [EPR18463] (ab191385) at 1/10000 dilution + Jurkat (Human T cell leukemia cell line from peripheral blood) cell lysate at 20 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

    Predicted band size: 39 kDa
    Observed band size: 42 kDa why is the actual band size different from the predicted?


    Exposure time: 30 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The observed MW and doublets are consistent with what has been described in the literature. Two bands run closely together as  doublets representing distinct isoforms; see UniProt annotation and PMID 24169447.

  • Western blot - Anti-SIRT6 antibody [EPR18463] (ab191385)
    Western blot - Anti-SIRT6 antibody [EPR18463] (ab191385)
    Anti-SIRT6 antibody [EPR18463] (ab191385) at 1/10000 dilution + NIH/3T3 (Mouse embryonic fibroblast cell line) cell lysate at 20 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

    Predicted band size: 39 kDa
    Observed band size: 39 kDa


    Exposure time: 30 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The observed MW and doublets are consistent with what has been described in the literature. Two bands run closely together as  doublets representing distinct isoforms; see UniProt annotation and PMID 24169447.

  • Western blot - Anti-SIRT6 antibody [EPR18463] (ab191385)
    Western blot - Anti-SIRT6 antibody [EPR18463] (ab191385)
    All lanes : Anti-SIRT6 antibody [EPR18463] (ab191385) at 1/5000 dilution

    Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) cytoplasmic lysate
    Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) nuclear lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 39 kDa
    Observed band size: 42 kDa why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

    SIRT6 is detected in nuclear fractions.

  • Western blot - Anti-SIRT6 antibody [EPR18463] (ab191385)
    Western blot - Anti-SIRT6 antibody [EPR18463] (ab191385)
    All lanes : Anti-SIRT6 antibody [EPR18463] (ab191385) at 1/2000 dilution

    Lane 1 : Rat brain lysate
    Lane 2 : Rat spleen lysate
    Lane 3 : C6 (Rat glial tumor cell line) cell lysate
    Lane 4 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) cell lysate
    Lane 5 : PC-12 (Rat adrenal gland pheochromocytoma cell line) cell lysate
    Lane 6 : NIH/3T3 (Mouse embryonic fibroblast cell line) cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

    Predicted band size: 39 kDa
    Observed band size: 39 kDa


    Exposure time: 30 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The observed MW is consistent with what has been described in the literature (PMID 24169447).

  • Immunocytochemistry/ Immunofluorescence - Anti-SIRT6 antibody [EPR18463] (ab191385)
    Immunocytochemistry/ Immunofluorescence - Anti-SIRT6 antibody [EPR18463] (ab191385)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling SIRT6 with ab191385 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on HeLa cell line. The nuclear counterstain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution (red).

    The negative controls are as follows:-

    -ve control 1: ab191385 at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution.

    -ve control 2: Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

  • Immunocytochemistry/ Immunofluorescence - Anti-SIRT6 antibody [EPR18463] (ab191385)
    Immunocytochemistry/ Immunofluorescence - Anti-SIRT6 antibody [EPR18463] (ab191385)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HCT 116 (Human colorectal carcinoma cell line) cells labeling SIRT6 with ab191385 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing nuclear staining on HCT 116 cell line.

    The nuclear counterstain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution (red).

    The negative controls are as follows:-

    -ve control 1: ab191385 at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution.

    -ve control 2: Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

  • Immunoprecipitation - Anti-SIRT6 antibody [EPR18463] (ab191385)
    Immunoprecipitation - Anti-SIRT6 antibody [EPR18463] (ab191385)

    SIRT6 was immunoprecipitated from 1 mg of Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate with ab191385 at 1/40 dilution.

    Western blot was performed from the immunoprecipitate using ab191385 at 1/2000 dilution.

    Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.

    Lane 1: Jurkat whole cell lysate 10µg (Input).

    Lane 2: ab191385 IP in Jurkat whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) IP instead of ab191385 in Jurkat whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 3 seconds.

    The observed MW and doublets are consistent with what has been described in the literature. Two bands run closely together as  doublets representing distinct isoforms; see UniProt annotation and PMID 24169447.

  • Anti-SIRT6 antibody [EPR18463] (ab191385)
    Anti-SIRT6 antibody [EPR18463] (ab191385)

文献 (10)

发表研究结果有使用 ab191385？请让我们知道，以便我们可以引用本数据表中的参考文章。

ab191385 被引用在 10 文献中.

  • Xu Y  et al. Sirt3 is a novel target to treat sepsis induced myocardial dysfunction by acetylated modulation of critical enzymes within cardiac tricarboxylic acid cycle. Pharmacol Res 159:104887 (2020). PubMed: 32526680
  • D'Onofrio N  et al. ROS-Mediated Apoptotic Cell Death of Human Colon Cancer LoVo Cells by Milk d-Valerobetaine. Sci Rep 10:8978 (2020). PubMed: 32488123
  • Wang T  et al. Sirt6 stabilizes atherosclerosis plaques by promoting macrophage autophagy and reducing contact with endothelial cells. Biochem Cell Biol N/A:N/A (2019). PubMed: 31063699
  • Zhang C  et al. SIRT6 regulates the proliferation and apoptosis of hepatocellular carcinoma via the ERK1/2 signaling pathway. Mol Med Rep 20:1575-1582 (2019). PubMed: 31257493
  • Zhang CW  et al. Long non-coding RNA PVT1 knockdown suppresses fibroblast-like synoviocyte inflammation and induces apoptosis in rheumatoid arthritis through demethylation of sirt6. J Biol Eng 13:60 (2019). PubMed: 31303891
  • Ji L  et al. Overexpression of Sirt6 promotes M2 macrophage transformation, alleviating renal injury in diabetic nephropathy. Int J Oncol 55:103-115 (2019). PubMed: 31115579
  • Rezazadeh S  et al. SIRT6 promotes transcription of a subset of NRF2 targets by mono-ADP-ribosylating BAF170. Nucleic Acids Res 47:7914-7928 (2019). PubMed: 31216030
  • Geng K  et al. Adjudin delays cellular senescence through Sirt3-mediated attenuation of ROS production. Int J Mol Med 42:3522-3529 (2018). PubMed: 30320336
  • Zhu B  et al. Downregulation of SIRT6 is associated with poor prognosis in patients with non-small cell lung cancer. J Int Med Res 46:1517-1527 (2018). PubMed: 29363378
  • Li N  et al. Downregulation of SIRT6 by miR-34c-5p is associated with poor prognosis and promotes colon cancer proliferation through inhibiting apoptosis via the JAK2/STAT3 signaling pathway. Int J Oncol N/A:N/A (2018). PubMed: 29512698

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