Overview

  • Product name
  • Description
    Rabbit polyclonal to SIRT2
  • Host species
    Rabbit
  • Specificity
    SIRT2 antibody (ab19388) detects SIRT2 and shows no cross-reactivity to other SIRT isoforms.
  • Tested applications
    Suitable for: WB, ICCmore details
  • Species reactivity
    Reacts with: Rat, Human
  • Immunogen

    Synthetic peptide corresponding to Human SIRT2 aa 377-389.
    Sequence:

    DEARTTEREKPQ

  • Positive control
    • SAOS2 transfected cell lysates, human chondrocyte lysates.
  • General notes


    The SIRT2 protein (also known as Sirtuin for Silent Mating Type Information 2-Homolog) is a NAD-dependent deacytylase (NDAC) that has been shown to control gene silencing, cell cycle, and DNA damage repair. It is believed that SIRT2 may act as a tumor suppressor in human gliomas and may also serve as a novel molecular marker for these cells. SIRT2 has also been shown to act as a redox sensor to help regulate muscle gene expression in response to food intake and exercise. SIRT2 acts in the phosphorylation cascade involving mitosis where SIRT2 is phosphorylated in late G2 phase, during M phase, and into cytokinesis.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer
    Preservative: 0.05% Sodium azide
  • Purity
    Whole antiserum
  • Primary antibody notes
    The SIRT2 protein (also known as Sirtuin for Silent Mating Type Information 2-Homolog) is a NAD-dependent deacytylase (NDAC) that has been shown to control gene silencing, cell cycle, and DNA damage repair. It is believed that SIRT2 may act as a tumor suppressor in human gliomas and may also serve as a novel molecular marker for these cells. SIRT2 has also been shown to act as a redox sensor to help regulate muscle gene expression in response to food intake and exercise. SIRT2 acts in the phosphorylation cascade involving mitosis where SIRT2 is phosphorylated in late G2 phase, during M phase, and into cytokinesis.
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab19388 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
WB 1/1000. Detects a band of approximately 48 kDa (predicted molecular weight: 43 kDa).
ICC 1/400.

Target

  • Function
    NAD-dependent protein deacetylase, which deacetylates the 'Lys-40' of alpha-tubulin. Involved in the control of mitotic exit in the cell cycle, probably via its role in the regulation of cytoskeleton.
  • Tissue specificity
    Widely expressed. Highly expressed in heart, brain and skeletal muscle, while it is weakly expressed in placenta and lung. Down-regulated in many gliomas suggesting that it may act as a tumor suppressor gene in human gliomas possibly through the regulation of microtubule network.
  • Sequence similarities
    Belongs to the sirtuin family.
    Contains 1 deacetylase sirtuin-type domain.
  • Developmental stage
    Peaks during mitosis. After mitosis, it is probably degraded by the 26S proteasome.
  • Post-translational
    modifications
    Phosphorylated at the G2/M transition of the cell cycle.
  • Cellular localization
    Cytoplasm > cytoskeleton. Colocalizes with microtubules.
  • Information by UniProt
  • Database links
  • Alternative names
    • FLJ35621 antibody
    • FLJ37491 antibody
    • NAD dependent deacetylase sirtuin 2 antibody
    • NAD-dependent deacetylase sirtuin-2 antibody
    • NAD-dependent protein deacetylase sirtuin-2 antibody
    • Regulatory protein SIR2 homolog 2 antibody
    • Silencing information regulator 2 like antibody
    • Silent information regulator 2 antibody
    • SIR2 antibody
    • SIR2 like protein 2 antibody
    • Sir2 related protein type 2 antibody
    • SIR2, S. cerevisiae, homolog-loke 2 antibody
    • SIR2-like protein 2 antibody
    • SIR2L antibody
    • SIR2L2 antibody
    • SIRT2 antibody
    • SIRT2_HUMAN antibody
    • Sirtuin (silent mating type information regulation 2 homolog) 2 (S.cerevisiae) antibody
    • Sirtuin 2 antibody
    • Sirtuin type 2 antibody
    see all

Images

  • Anti-SIRT2 antibody (ab19388) at 1/1000 dilution
    Predicted band size: 43 kDa



    By Western blot, this antibody detects an ~48 kDa protein representing SIRT2 from transfected SAOS2 cells.
  • Immunofluorescence staining of SIRT2 in transfected SAOS2 cells using ab19388.

References

This product has been referenced in:
  • Lara E  et al. Salermide, a Sirtuin inhibitor with a strong cancer-specific proapoptotic effect. Oncogene 28:781-91 (2009). WB ; Human . Read more (PubMed: 19060927) »
  • Lovato L  et al. Transketolase and CNPase I are specifically recognized by IgG autoantibodies in multiple sclerosis patients. Mol Cell Proteomics : (2008). WB ; Human . Read more (PubMed: 18676363) »
See all 2 Publications for this product

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

Question
Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement, the rabbit monoclonal ab134171.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

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Question

BATCH NUMBER 156492 ORDER NUMBER 013816 DESCRIPTION OF THE PROBLEM The expected band of SIRT-2 in Western Blot should be about 48 kd - we got 7 bands with different sizes (range from 33-97 kd) and we are not sure which one is the specific SIRT-2 band. SAMPLE hippocampal lysates (brain tissure) PRIMARY ANTIBODY SIRT-2 (ab19388) rabbit polyclonal AB lot#156492 Abcam diluent: wash buffer (Tween-20 in 1x PBS/azide)/blocking buffer (1:1) incubate: 1h RT dilution: 1:1000 wash: 3 x 10min. in blocking buffer DETECTION METHOD Chemiluminescent Substrate: Nitroblock II / CDP Star Tropix ANTIBODY STORAGE CONDITIONS -20C SAMPLE PREPARATION RIPA buffer + Protease inhibitors (no Sonication used) add loading buffer and boiled for 5 min. then froze samples Date of lysate preparations and storage temp: 10/05/2005 -20C AMOUNT OF PROTEIN LOADED 20ug/well ELECTROPHORESIS/GEL CONDITIONS 10 % acrylamide/SDS gel TRANSFER AND BLOCKING CONDITIONS semidry blot to PVDF membrane (Millipore) 20V limit 0.1A 150min (transfer time) blocking buffer: I-block (Tropix), Tween-20 in 1x PBS/azide incubate: 1h at roomtemperature (RT) SECONDARY ANTIBODY (goat) anti-rabbit conjugated to Alkal.Phosph. 2nd AB #6520 Tropix diluent: wash buffer (Tween-20 in 1x PBS/azide)/blocking buffer (1:1) incubate: 1h RT dilution: 1:10000 wash: 3 x 10min. in blocking buffer HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 1x HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes ADDITIONAL NOTES attached you will find the result of our Western Blot with SIRT-2 Thank you very much for your help.

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Answer

I'm sorry to hear you are experiencing problems with ab19388. Many thanks for providing the image of your blot and your detailed protocol, it is very useful for me to understand your problem. I would like to recommend some changes to your protocol to increase the signal specific to SIRT2 and to decrease the background too. I am not sure how high levels of SIRT2 are in the adult brain so I would recommend to run a positive control along your samples, such as mouse 3T3 lysate. Is the species you are using human or rat? I am also concerned that your samples date from last October and were stored at -20C if they were stored in lysis buffer. If they were stored after boiling in loading buffer they should be fine. Were there plenty of fresh protease inhibitors at the time of lysis, could they have gone off? I would recommend to incubate the membrane at 4C overnight and to try a range of dilutions (you may be able to dilute more by incubating longer), this will give a slow but targeted binding. To improve on the background on the membrane I would recommend to try blocking 1hr only in BSA (5% in TBST) and to incubate the antibodies in TBST only, at 4C. We recommend to use HRP conjugated secondary antibodies and ECl+ detection systems, so if you have those available it may be worth trying too, as they give less background. Please also wash the membrane in TBST only, this should help too. I hope the above recommendations will help you, please do not hesitate to contact me again if you need further assistance, ***An error on the datasheet regarding testing in mouse samples has since been detected and the customer was refunded her order. Mouse samples have not been tested***

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Answer

Thank you for contacting us for technical support. The fact the customer is having the correct results with over-expressed protein reassures us that the antibody works and that in Hek293T cell lysate the protein levels are much lower. We have not had any complaints about these antibodies and they work very well in the tested positive controls. I would recommend running the positive controls detailed on the datasheets: ab10140: human kidney cell lysate, U937 lysate ab19388: 3T3 cell lysate ab13860: no cell line has been tested- please try human testis lysate ab13697: no cell line has been tested- please try human intestine lysate If you require further assistance can you please ask your customer to fill in our protocol questionnaires and we can look at each antibody condition; typically please check your customer incubates at the recommended dilution in TBST overnight (he/she may need to use more concentrated antibody as this depends on the tissue), please check that degradation of the protein is not occurring and other protocol details which may make a difference,

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Answer

Thank you for your email, and I'm replying on behalf of Sarah who is currently away from the office. I'm sorry to hear that ab19388 is still giving you difficulty and I can certainly arrange for a free of charge replacement vial to be sent to you. As you ordered via one of distributors, Biozol, please notify them of the situation (forward my email to them) and they will send you the replacement. Please let us know how the replacement works out for you and if you have any additional questions.

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Question

BATCH NUMBER 107843 ORDER NUMBER -- NOT SPECIFIED -- DESCRIPTION OF THE PROBLEM Multiple bands SAMPLE Cell extract of A549 lung cancer cells, AsPC-1 pancreas carcinoma cells and HCT116 colon carconima cells PRIMARY ANTIBODY ab19388, 1:1000 and 1:2000 in blocking reagent incubation over night at 4?C washing at room temperature with TBS-T, 3x 15min DETECTION METHOD ECL (Amersham) ANTIBODY STORAGE CONDITIONS antibody arrived at room temperature and was stored at -20?C immediately SAMPLE PREPARATION RIPA buffer + protease inhibitors (complete Mini tablets, Roche) + Vanadate + PMSF + NaF samples were mixed with L?mmli buffer and heated to 95?C for 5min AMOUNT OF PROTEIN LOADED 50?g total protein ELECTROPHORESIS/GEL CONDITIONS SDS-PAGE electrophoresis 10% gel TRANSFER AND BLOCKING CONDITIONS Transfer buffer: 25mM Tris, 192mM Glycine transfer 0,1A/gel, 90min blocking with 3% BSA in TBS-T SECONDARY ANTIBODY goat-anti-rabbit (Biomol), 1:5000 in blocking reagent incubation at room temperature for 1h washing at room temperature with TBS-T, 3x 15min HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 2 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? antibody dilution ADDITIONAL NOTES the same samples were used for detection of HDAC6 expression using the same secondary antibody giving one distinct band antibody was ordered via Biozol (oder number: 43045453491) I am on a holiday from 26. August to 16. September for futher questions please contact Heike Wieland: heike.wieland@altanapharma.com

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Answer

Thank you for contacting us for technical support and for taking the time to provide all your protocol details, this was really useful to understand your problem. I suspect the problem is due to the blocking agent present in your antibody dilution buffer. I would recommend trying: -5%BSA 1hr then incubating the primary and secondary in TBST only -5% milk 1hr then incubating the primary and secondary in TBST only This will give sufficient blocking to prevent non specific binding but we know that too much blocking can also cause non specific binding, so a balance is essential. The rest of your protocol looks really good so I think you will have much better results with this change (though you can also try to load a little less sample and dilute more your secondary and do more washing steps too) (also make sure the secondary works well with other primary antibodies). Please let me know if you still experience a problem and I can offer you a replacement vial or refund if you purchased the antibody in the last 90days

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