Overview

  • Product name
  • Description
    Goat polyclonal to SIRT4
  • Host species
    Goat
  • Tested applications
    Suitable for: WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Rat, Human
    Predicted to work with: Mouse, Cow, Dog, Pig
  • Immunogen

    Synthetic peptide:

    SRCGELLPLIDPC

    , corresponding to C terminal amino acids 302-314 of Human SIRT4.

  • Positive control
    • Human kidney and U937 lysate. Humsn skeletal muscle tissue.
  • General notes


    This gene encodes a member of the sirtuin family of proteins, homologs to the yeast Sir2 protein. Members of the sirtuin family are characterized by a sirtuin core domain and grouped into four classes. The functions of human sirtuins have not yet been determined; however, yeast sirtuin proteins are known to regulate epigenetic gene silencing and suppress recombination of rDNA. Studies suggest that the human sirtuins may function as intracellular regulatory proteins with mono-ADP-ribosyltransferase activity. The protein encoded by this gene is included in class IV of the sirtuin family.

Properties

Applications

Our Abpromise guarantee covers the use of ab10140 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 Use a concentration of 1 - 3 µg/ml. Detects a band of approximately 38 kDa (predicted molecular weight: 36 kDa).Can be blocked with Human SIRT4 peptide (ab23185).
IHC-P Use a concentration of 2 - 3 µg/ml. Steamed antigen retrieval with citrate buffer pH 6 is recommended.

Target

  • Function
    Acts as NAD-dependent protein lipoamidase, ADP-ribosyl transferase and deacetylase. Catalyzes more efficiently removal of lipoyl- and biotinyl- than acetyl-lysine modifications. Inhibits the pyruvate dehydrogenase complex (PDH) activity via the enzymatic hydrolysis of the lipoamide cofactor from the E2 component, DLAT, in a phosphorylation-independent manner (PubMed:25525879). Catalyzes the transfer of ADP-ribosyl groups onto target proteins, including mitochondrial GLUD1, inhibiting GLUD1 enzyme activity. Acts as a negative regulator of mitochondrial glutamine metabolism by mediating mono ADP-ribosylation of GLUD1: expressed in response to DNA damage and negatively regulates anaplerosis by inhibiting GLUD1, leading to block metabolism of glutamine into tricarboxylic acid cycle and promoting cell cycle arrest (PubMed:16959573, PubMed:17715127). In response to mTORC1 signal, SIRT4 expression is repressed, promoting anaplerosis and cell proliferation. Acts as a tumor suppressor (PubMed:23562301, PubMed:23663782). Also acts as a NAD-dependent protein deacetylase: mediates deacetylation of 'Lys-471' of MLYCD, inhibiting its activity, thereby acting as a regulator of lipid homeostasis (By similarity). Controls fatty acid oxidation by inhibiting PPARA transcriptional activation. Impairs SIRT1:PPARA interaction probably through the regulation of NAD(+) levels (PubMed:24043310). Down-regulates insulin secretion.
  • Tissue specificity
    Detected in vascular smooth muscle and striated muscle. Detected in insulin-producing beta-cells in pancreas islets of Langerhans (at protein level). Widely expressed. Weakly expressed in leukocytes and fetal thymus.
  • Sequence similarities
    Belongs to the sirtuin family. Class II subfamily.
    Contains 1 deacetylase sirtuin-type domain.
  • Cellular localization
    Mitochondrion matrix.
  • Information by UniProt
  • Database links
  • Alternative names
    • MGC130046 antibody
    • MGC130047 antibody
    • MGC57437 antibody
    • NAD dependent deacetylase sirtuin 4 antibody
    • NAD dependent protein deacetylase sirtuin 4 antibody
    • NAD-dependent ADP-ribosyltransferase sirtuin-4 antibody
    • NAD-dependent protein deacetylase sirtuin-4 antibody
    • NAD-dependent protein lipoamidase sirtuin-4, mitochondrial antibody
    • Regulatory protein SIR2 homolog 4 antibody
    • Silent mating type information regulation 2 homolog antibody
    • Sir 2 like 4 antibody
    • SIR 2 like protein 4 antibody
    • Sir2 like 4 antibody
    • SIR2 like protein 4 antibody
    • SIR2-like protein 4 antibody
    • SIR2L 4 antibody
    • SIR2L4 antibody
    • SIR4_HUMAN antibody
    • SIRT 4 antibody
    • Sirt4 antibody
    • Sirtuin (silent mating type information regulation 2 homolog) 4 antibody
    • Sirtuin (silent mating type information regulation 2 homolog) 4 (S. cerevisiae) antibody
    • Sirtuin 4 antibody
    • Sirtuin type 4 antibody
    • Sirtuin-4 antibody
    • Sirtuin4 antibody
    see all

Images

  • ab10140 at 2.5µg/ml staining SIRT4 in human skeletal muscle tissue section by Immunohistochemistry (Formalin/ PFA fixed paraffin-embedded tissue sections). Tissue underwent antigen retrieval in steam in citrate buffer pH 6.0. AP staining procedure was used for detection.

  • Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human pancreas tissue, staining SIRT4 with ab10140 at 2 µg/ml. Antigen retrieval was performed by heat mediation in a citrate buffer (pH 6).
  • Anti-SIRT4 antibody (ab10140) at 0.05 µg/ml + Human kidney lysate in RIPA buffer at 35 µg

    Developed using the ECL technique.

    Predicted band size: 36 kDa
    Observed band size: 37 kDa
    why is the actual band size different from the predicted?

  • All lanes : Anti-SIRT4 antibody (ab10140) at 3 µg/ml

    Lane 1 : MOCK-transfected HEK293 cell lysate
    Lane 2 : SIRT4-transfected HEK293 cell lysate

    Predicted band size: 36 kDa
    Observed band size: 38 kDa why is the actual band size different from the predicted?

References

This product has been referenced in:
  • Zhu Y  et al. Knockout of SIRT4 decreases chemosensitivity to 5-FU in colorectal cancer cells. Oncol Lett 16:1675-1681 (2018). Read more (PubMed: 30008852) »
  • Luo H  et al. Expression of Sirtuins in the Retinal Neurons of Mice, Rats, and Humans. Front Aging Neurosci 9:366 (2017). WB, IHC-P ; Mouse, Rat, Human . Read more (PubMed: 29249955) »
See all 8 Publications for this product

Customer reviews and Q&As

1-8 of 8 Abreviews or Q&A

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Brain (coronal sections))
Specification
Brain (coronal sections)
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.3% Triton X-100
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Apr 15 2013

Answer

That is correct. The antibody is goat immunoglobulin, purified from goat serum. The immunogen is derived from human Sirt4, but the antibody itself is only goat IgG. If you or your customer still have any questions, please contact us.

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Answer

Thanks again for contacting us with your question, and for your patience. The peptide for ab49173 is now available on our website as ab123888- https://www.abcam.com/SIRT5-peptide-ab123888.html Unfortunately the peptide for ab78977 is not available for sale. I am sorry that we don't have all of the products you are looking for, but please let me know if you have any questions or if there is anything else that we can do for you.  

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Answer

Thank you for contacting us with your question. We do have the blocking peptide for the SIRT5 antibody ab49173, but it isn't on our website at this point so I'm working to have this added. I'll have to check with the lab about the peptide for the SIRT7 antibody ab78977, and I'll get back to you once I have more information about its availability. Unfortunately, the peptide for the SIRT1 antibody ab32441 is not available for sale. I will be in touch once I hear about the ab49173 and ab78977 peptides, but please let me know if you have any questions in the meantime.

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Answer

Thank you for kindly confirming these details as this enables us to closely monitor the quality of our products. We would be pleased to arrange a replacement product free of charge. I am forwarding these details on to our Distributor’s Team who would be happy to organize this. Distributor name:Bita Lifescience  Original order number:xxxabID of original product: 1 X vial ab78982 Original batch number:1062565  abID of replacement product: ab10140 CCEID:xxxThank you for your help and cooperation. Please do not hesitate to contact us if you need anything further.

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Answer

Thank you very much for confirming these details. I would be pleased to replace the antibody with an Sirt4 antibody. If the customer is interested in an antibody for IHC-P, I agree that ab10140, ab105039 and ab115517 are suitable. As mentioned on the datasheet all of these require heat-mediated antigen retrieval. This should preferably performed with citrate buffer at pH6, however, might require optimisation according to the experimental setting, especially in the case of staining PFA-perfused mouse brain sections. As for the Sirt5 antibody ab78982, we do not know if these Sirt4 antibodies would work with a protocol for free floating sections and thus the protocol might require optimisation. Also, these antibodies have so far only been tested with human (and rat) samples, however, due to a high sequence homology we expect them to cross-react with murine Sirt4 as well. Therefore, if the customer is happy to try ab10140 (or one of the others) despite these uncertainties, I would be pleased to arrange for a replacement. Otherwise, I could issue a credit note or refund if preferred. Thank you for your cooperation. I look forward to hearing from you with details of how your customer would like to proceed.

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Answer

Thank you for your customer's protocol details, I enclose below my recommendations to increase the signal, these are very similar to those given 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 kidney and U937 lysates (the customer's RIPA buffer is a good choice). -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 I would therefore recommend blocking the membrane in 5% BSA (30min-1hrs) and incubating the antibody in TBST only. -please try 1:300 dilution with overnight incubation. We have used 1ug/ml (i.e 1:500) in our positive control but this needs optimisation between samples. - 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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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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