Overview

  • Product name
  • Description
    Rabbit polyclonal to SIRT5
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide:

    CLKMARPSSSMADFRKF

    , corresponding to amino acids 30-46 of Human SIRT5.

  • Positive control
    • Human rat or mouse intestine tissue lysate.

Properties

Applications

Our Abpromise guarantee covers the use of ab13697 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 - 1/2000. Detects a band of approximately 36 kDa.

Target

  • Function
    NAD-dependent lysine demalonylase, desuccinylase and deglutarylase that specifically removes malonyl, succinyl and glutaryl groups on target proteins (PubMed:21908771, PubMed:22076378, PubMed:24703693). Activates CPS1 and contributes to the regulation of blood ammonia levels during prolonged fasting: acts by mediating desuccinylation and deglutarylation of CPS1, thereby increasing CPS1 activity in response to elevated NAD levels during fasting (PubMed:22076378, PubMed:24703693). Activates SOD1 by mediating its desuccinylation, leading to reduced reactive oxygen species (PubMed:24140062). Modulates ketogenesis through the desuccinylation and activation of HMGCS2 (By similarity). Has weak NAD-dependent protein deacetylase activity; however this activity may not be physiologically relevant in vivo. Can deacetylate cytochrome c (CYCS) and a number of other proteins in vitro such as UOX.
  • Tissue specificity
    Widely expressed.
  • Sequence similarities
    Belongs to the sirtuin family. Class III subfamily.
    Contains 1 deacetylase sirtuin-type domain.
  • Domain
    In contrast to class I sirtuins, class III sirtuins have only weak deacetylase activity. Difference in substrate specificity is probably due to a larger hydrophobic pocket with 2 residues (Tyr-102 and Arg-105) that bind to malonylated and succinylated substrates and define the specificity (PubMed:22076378).
  • Cellular localization
    Mitochondrion; Cytoplasm. Mitochondrion and Mitochondrion matrix. Mitochondrion intermembrane space. Cytoplasm, cytosol. Nucleus. Mainly mitochondrial. Also present extramitochondrially: a fraction is present in the cytosol and very small amounts are also detected in the nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • NAD dependent deacetylase sirtuin 5 antibody
    • NAD dependent lysine demalonylase and desuccinylase sirtuin 5 mitochondrial antibody
    • NAD dependent protein deacylase sirtuin 5 mitochondrial antibody
    • NAD-dependent protein deacylase sirtuin-5, mitochondrial antibody
    • Regulatory protein SIR2 homolog 5 antibody
    • Silent mating type information regulation 2 S.cerevisiae homolog 5 antibody
    • Sir2 like 5 antibody
    • SIR2-like protein 5 antibody
    • SIR2L5 antibody
    • SIR5_HUMAN antibody
    • Sirt5 antibody
    • Sirtuin 5 antibody
    • Sirtuin type 5 antibody
    see all

Images

  • Anti-SIRT5 antibody (ab13697) at 1/1000 dilution + Human intestine lysate


    Western blot analysis of SIRT5 using ab13697 at 1:1000 dilution against 20 µg of human intestine lysate.

  • All lanes : Anti-SIRT5 antibody (ab13697) at 2 µg/ml

    Lane 1 : Human intestine cell lysate
    Lane 2 : Mouse intestine cell lysate
    Lane 3 : Rat intestine cell lysate

    Observed band size: 36 kDa
    why is the actual band size different from the predicted?

References

This product has been referenced in:
  • Fukushima A  et al. Acetylation contributes to hypertrophy-caused maturational delay of cardiac energy metabolism. JCI Insight 3:N/A (2018). Read more (PubMed: 29769443) »
See 1 Publication for this product

Customer reviews and Q&As

1-9 of 9 Abreviews or Q&A

Answer

Thank you for your reply.

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 of ab108968 with the order number XXX.

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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Answer

According to our records, ab13697 was proving difficult to use in WB and we were in contact in order to help resolve the issue.

Looking at our correspondence, it appears that we are awaiting more details in order to help us better understand the difficulties experienced. If the requested information has already been sent, it appears that it did not reach our Scientific Support team and we apologize for this inconvenience. In this case we would like to ask for the information again so that we can reach a resolution.

If the issue has already been settled, please let us know so that we can be assured that the problem has been solved to your satisfaction and update our records.
We wish you the best of luck with your research and look forward to a reply.

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Answer

Thank you for your honest feedback. I agree that the vial of ab13697 you received is not specific for SIRT5. As per our Abpromise guarantee, if you have received this product within the last 6 months, I am happy to offer a free replacement of any primary antibody, credit or refund.
In order to process your request, please include the PO number or Abcam order reference number associated with the purchase of ab13697 in your reply.
Thanks!

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Application
Western blot
Sample
Mouse Tissue lysate - other (Liver mitochondrial extract)
Loading amount
20 µg
Specification
Liver mitochondrial extract
Gel Running Conditions
Reduced Denaturing (10 %)
Blocking step
LiCOR blocking buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C

Dr. Michelle Green

Verified customer

Submitted Aug 08 2012

Answer

Thank you for contacting Abcam regarding ab13697.

I am sorry that this antibody is not working as specified on the datasheet. I am happy to offer a replacement or credit for this product.

Unfortunately, I am not able to send samples of all of our SIRT5 antibodies, however I would like to suggest some alternatives and I am happy to offer one as a free of charge replacement. In particular, I think ab108968 would be a great alternative.

Please let me know if you are interested in receiving this product. Please provide an order number for ab13697 in your reply.

Finally, if possible, I would like to get the protocol details for ab13697. I will forward this on to the lab so that we may investigate this issue further.

I look forward to your reply so that I may assist you further. Please do not hesitate to contact me if you have any additional questions or concerns.

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Answer

Thank you for contacting us.

Your credit note ID is xxx.


As per our order system, both ab13697 and ab90845 have already been refunded in xxx with credit note ID xxx.


The current credit note is for ab125881.

I am sorry that this antibody did not perform as stated on the datasheet. If payment has already been made on the original order and you wish to receive a refund, please ask your purchasing department to contact our accounting department so that we may gather the correct information needed for the refund. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used.

Our accounting department can be contacted by email at us.credits@abcam.com or by telephone using the information at the Contact Us link in the top right corner of our website. Please refer to the credit note ID in any correspondence with our accounting department.

The credit note ID is for your reference only and does not automatically guarantee the credit.

I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service, should you require further expert advice.

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Question
Answer

Thank you for your call today and for letting us know about the trouble with these antibodies.

Your credit note ID is ***.

I am sorry that these antibodies did not perform as stated on the datasheet. If payment has already been made on the original order and you wish to receive a refund, please ask your purchasing department to contact our accounting department so that we may gather the correct information needed for the refund. Our accounting department can be contacted by email at us.credits@abcam.com or by telephone at 888-772-2226. Please refer to the credit note ID in any correspondence with our accounting department.

I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service, should you require further assistance or advice.

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Question

Lot no: 137348 Methods: Westernblot Abcam catalog: ab13697 1. Antibody Storage Conditions (temperature/reconstitution etc): -20’C, aliquot 2. Description of the problem (high background, wrong band size, more bands, no band etc.) : high background, wrong band size, more bands 3. Sample (Species/Cell extract/Nuclear extract/Purified protein/Recombinant protein etc.) : HEK293T whole cell extract 4. Sample preparation (Buffer/Protease inhibitors/Heating sample etc.) : RIPA buffer with protease inhibitors, heating sample 5. Amount of protein loaded : 50 ug 6. Electrophoresis/Gel conditions (Reducing or Non-reducing gel, % of the gel etc.) : reducing gel, 10 ~ 15 % 7. Transfer and blocking conditions (Buffer/Time period, Blocking agent etc.) : Tris/Glycine/MeOH/0.1%SDS transfer buffer (deeply stained by PonceauS, other antibodies were worked well), blocking agent : 3 % non-fat milk in TBST 8. Primary antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, wash step) : abcam/Rabbit/3 % non-fat milk in TBST /1:500~1:1000/2hours~overnight incubation, 30 min. washing 9. Secondary antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, wash step) : Jackson/Goat/3 % non-fat milk in TBST /1:1000~1:5000/1hour incubation, 30 min.~1hour washing 10. Detection method (ECL, ECL Plus etc.) : ECL 11. Positive and negative controls used (Please specify) : Purified protein, siRNA 12. How many times have you tried the Western? : 3 times 13. Have you run a “No Primary” control? (Yes or No) : Yes 14. Do you obtain the same results every time? (Yes or No) : Yes 15. What steps have you altered? : loading quantity 16. Additional Notes

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Answer

Thank you for providing more details of your customer's protocol, below are my recommendations to increase the signal (those are very similar to my recommendations for the other antibodies the customer is having problems with): -as discussed before 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. I would recommend running the positive control of Human rat or mouse intestine tissue lysate. -check the protease inhibitors in the lysis buffer are fresh and not old and that samples are kept on ice at all times- can I also check that the samples are left to lyse for 1hr before spinning in the centrifuge (to give time for the NP40 to permeabilise and lyse the cells)? -milk can prevent binding of the antibody to the protein and also give high background unfortunately I would therefore recommend blocking the membrane in 5% BSA (30min-1hrs) and incubating the antibody in TBST only. - use Ecl+ or even more sensitive kits (e.g. West Pico, West dura pierce kits) as ECL is not very sensitive -check the secondary antibody is not too old and not loosing its sensitivity, if possible by trying a different secondary antibody. I hope these suggestions will help,

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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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