Recombinant 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
Related conjugates and formulations
Overview
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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.
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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.
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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
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
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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...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR18463 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
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 |
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ICC/IF |
1/1000.
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IP |
1/40.
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WB | (1) |
1/2000. Detects a band of approximately 42 kDa (predicted molecular weight: 39 kDa).
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Notes |
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ICC/IF
1/1000. |
IP
1/40. |
WB
1/2000. Detects a band of approximately 42 kDa (predicted molecular weight: 39 kDa). |
Target
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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. - Information by UniProt
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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
see all -
Alternative names
- 2810449N18Rik antibody
- AI043036 antibody
- Mono ADP ribosyltransferase sirtuin 6 antibody
see all
Images
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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.
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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.
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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 minutesBlocking/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.
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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 secondsBlocking/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.
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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 secondsBlocking/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.
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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 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
SIRT6 is detected in nuclear fractions.
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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 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
The observed MW is consistent with what has been described in the literature (PMID 24169447).
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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.
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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.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (16)
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