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    products/primary-antibodies/sirt6-antibody-epr18463-ab191385.pdf

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Epigenetics and Nuclear Signaling Nuclear Signaling Pathways NFkB pathway
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-SIRT6 antibody [EPR18463] (ab191385)

  • Datasheet
  • SDS
  • Certificate of Compliance
Reviews (2) Submit a question References (16)

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

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

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 647 Carrier Free

You may also be interested in

Primary
Product image
Anti-Brd4 antibody [EPR5150(2)] (ab128874)
Knockout
Product image
Human SIRT6 knockout HeLa cell line (ab265054)
Assay
Product image
SIRT6 Inhibitor Screening Kit (Fluorometric) (ab283376)

View more associated products

Overview

  • Product name

    Anti-SIRT6 antibody [EPR18463]
    See all SIRT6 primary antibodies
  • Description

    Rabbit monoclonal [EPR18463] to SIRT6
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, IP, WBmore details
    Unsuitable for: ChIP-seq
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • 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.
  • General notes

    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.

Properties

  • Form

    Liquid
  • Storage instructions

    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.
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR18463
  • Isotype

    IgG
  • Research areas

    • Epigenetics and Nuclear Signaling
    • Nuclear Signaling Pathways
    • NFkB pathway
    • Epigenetics and Nuclear Signaling
    • Chromatin Modifying Enzymes
    • Acetylation
    • HDACs
    • Class III / Sir2 class
    • Cancer
    • Signal transduction
    • Nuclear signaling
    • NFkB pathway
    • Cardiovascular
    • Atherosclerosis
    • Diabetes associated
    • Cardiovascular
    • Atherosclerosis
    • Apoptosis
    • Metabolism
    • Types of disease
    • Obesity

Associated products

  • Alternative Versions

    • Alexa Fluor® 647 Anti-SIRT6 antibody [EPR18463] (ab225363)
    • Alexa Fluor® 488 Anti-SIRT6 antibody [EPR18463] (ab225364)
    • Anti-SIRT6 antibody [EPR18463] - BSA and Azide free (ab236024)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human SIRT6 knockout HeLa cell line (ab265054)
  • KO cell lysates

    • Human SIRT6 knockout HeLa cell lysate (ab257673)
  • Recombinant Protein

    • Recombinant human SIRT6 protein (ab104030)

Applications

The Abpromise guarantee

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.

Application Abreviews Notes
ICC/IF
1/1000.
IP
1/40.
WB (1)
1/2000. Detects a band of approximately 42 kDa (predicted molecular weight: 39 kDa).
Notes
ICC/IF
1/1000.
IP
1/40.
WB
1/2000. Detects a band of approximately 42 kDa (predicted molecular weight: 39 kDa).
Application notes
Is unsuitable for ChIP-seq.

Target

  • Function

    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.
  • Sequence similarities

    Belongs to the sirtuin family. Class IV subfamily.
    Contains 1 deacetylase sirtuin-type domain.
  • Cellular localization

    Nucleus, nucleoplasm. Predominantly nuclear. Associated with telomeric heterochromatin regions.
  • Target information above from: UniProt accession Q8N6T7 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 51548 Human
    • Entrez Gene: 50721 Mouse
    • Entrez Gene: 299638 Rat
    • Omim: 606211 Human
    • SwissProt: Q8N6T7 Human
    • SwissProt: P59941 Mouse
    • Unigene: 423756 Human
    • Unigene: 131825 Mouse
    • Unigene: 25643 Mouse
    see all
  • Alternative names

    • 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

Images

  • 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.

     

     

  • 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.

  • 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 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)

Protocols

  • Western blot protocols
  • Immunoprecipitation protocols
  • Immunocytochemistry & immunofluorescence protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

Certificate of Compliance

To download a Certificate of Compliance, please enter your Lot number below:

References (16)

Publishing research using ab191385? Please let us know so that we can cite the reference in this datasheet.

ab191385 has been referenced in 16 publications.

  • Huang Y  et al. SIRT6-specific inhibitor OSS-128167 exacerbates diabetic cardiomyopathy by aggravating inflammation and oxidative stress. Mol Med Rep 23:N/A (2021). PubMed: 33760202
  • Gao F  et al. USP10 alleviates sepsis-induced acute kidney injury by regulating Sirt6-mediated Nrf2/ARE signaling pathway. J Inflamm (Lond) 18:25 (2021). PubMed: 34412625
  • 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
  • Cao Z  et al. Cumulus cell-derived and maternal SIRT6 differentially regulates porcine oocyte meiotic maturation. Theriogenology 142:158-168 (2020). PubMed: 31593883
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
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1-2 of 2 Abreviews or Q&A

Western blot abreview for Anti-SIRT6 antibody [EPR18463]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Pig Tissue lysate - whole (porcine endometrial explants)
Gel Running Conditions
Reduced Denaturing (10% TGX Stain-Free Gel)
Loading amount
30 µg
Specification
porcine endometrial explants
Blocking step
Milk as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 5% · Temperature: RT°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Apr 01 2022

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-SIRT6 antibody [EPR18463]

Good
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Pig Tissue sections (uterus)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate buffer pH 6
Permeabilization
No
Specification
uterus
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: RT°C
Fixative
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Apr 01 2022

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