Overview

  • Product name
  • Description
    Rabbit polyclonal to UCP3
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IHC-FoFr, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide corresponding to Human UCP3 aa 254-267 conjugated to keyhole limpet haemocyanin.
    Database link: P55916

  • General notes
    Storage in frost-free freezers is also not recommended. If slight turbidity occurs upon prolonged storage, clarify the solution by centrifugation before use. Working dilution samples should be discarded if not used within 12 hours.

Properties

Applications

Our Abpromise guarantee covers the use of ab10985 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. Predicted molecular weight: 34 kDa. Predicted molecular weight migrates at ~50 kDa in positive control lysate. (Protein is glycosylated)
IHC-FoFr Use at an assay dependent concentration.
IHC-P 1/3000.

Target

  • Function
    UCP are mitochondrial transporter proteins that create proton leaks across the inner mitochondrial membrane, thus uncoupling oxidative phosphorylation. As a result, energy is dissipated in the form of heat. May play a role in the modulation of tissue respiratory control. Participates in thermogenesis and energy balance.
  • Tissue specificity
    Only in skeletal muscle and heart. Is more expressed in glycolytic than in oxidative skeletal muscles.
  • Involvement in disease
    Defects in UCP3 may be involved in obesity (OBESITY) [MIM:601665]. It is a condition characterized by an increase of body weight beyond the limitation of skeletal and physical requirements, as the result of excessive accumulation of body fat.
  • Sequence similarities
    Belongs to the mitochondrial carrier family.
    Contains 3 Solcar repeats.
  • Cellular localization
    Mitochondrion inner membrane.
  • Information by UniProt
  • Database links
  • Form
    UCP3 is preferentially expressed in muscle
  • Alternative names
    • Mitochondrial uncoupling protein 3 antibody
    • SLC25A9 antibody
    • Solute carrier family 25 member 9 antibody
    • UCP 3 antibody
    • UCP3 antibody
    • UCP3_HUMAN antibody
    • Uncoupling protein 3 antibody
    • Uncoupling protein 3 mitochondrial proton carrier antibody
    see all

Images

  • ab10985 (2µg/ml) staining UCP3 in human skeletal muscle using an automated system (DAKO Autostainer Plus). Using this protocol there is strong cytoplasmic staining of myocytes.
    Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.

References

This product has been referenced in:
  • Wang YN  et al. Low-Dose 4-Hydroxy-2-Nonenal (HNE) Reperfusion Therapy Displays Cardioprotective Effects in Mice After Myocardial Infarction That Are Abrogated by Genipin. Med Sci Monit 24:3702-3709 (2018). Read more (PubMed: 29858912) »
  • Trewin AJ  et al. Acute exercise alters skeletal muscle mitochondrial respiration and H2O2 emission in response to hyperinsulinemic-euglycemic clamp in middle-aged obese men. PLoS One 12:e0188421 (2017). Read more (PubMed: 29161316) »
See all 7 Publications for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Answer

Thank you for sending on these details. The UCP3 protein that this antibody recognises undergoes a lot of post transcriptional modification events. The bands seen in the western blot are probably from the heavier glycosylated forms of the protein, which appear to be slightly different in muscle tissue and liver. Positive controls run using this antibody have shown bands at approximately 50kD. If the customer would like to be sure of this, they can de-glycosylate the protein before denaturing and running on the gel. (there are kits available for this if they do a search.) The bands should then run at approximately 34kD. I hope this helps. Please get back to me if there are any more questions.

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Question
Answer

Thank you for contacting us and taking the time to fill out the questionaire. I am sorry you have had trouble using this rabbit polyclonal anti-UCP3 antibody (ab10985) I have had a look at your procedure and have come up with the following suggestions to optimise your protocol which I hope will solve the problem. 1. Sample buffer Multiple band and bands of incorrect size can occur when the protein polymerises eg to dimers (which will run down the gel at a different speed). This can be prevented by ensuring adequate denaturing. Check that your loading buffer contains SDS for denaturing the protein and you could try heating for 10 minutes rather than 5 minutes. The samples should then be placed immediately on ice and run on the gel as soon as possible before the protein starts to form multimers again. 2. You have not mentioned how long you block for. It may be worth optimising your blocking step. You can try blocking overnight, or blocking at 4oC for shorter period of time. 3. The non specific binding could be due to excess of antibody. If this is the case,the suggested dilution on the datasheet is 1:1000, it would be better to titrate the antibody more dilute than this rather than less dilute as you have been doing. Your secondary antibody may also need titrating to prevent non specific binding. 4. I would not recommend you load any more than 100ug of protein per lane as this is fairly high. This will increase risk of multimers forming. 5. For your positive control, you could use a stronger lysing agent such as RIPA buffer (50mm TRis HCl, ph8, 150mM NaCl, 1% NP-40, 0.5%sodium deoxycholate, 0.1%SDS) and check the protien concentration in your tissue sample preparation to make sure there is enough protein. I hope this helps. Good luck with your experiment, and please let me know how you get on. If you have any more questions, or need any more advice, please contact me again. With thanks.

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Mouse Tissue sections (Skeletal muscle (mouse))
Specification
Skeletal muscle (mouse)
Fixative
Formaldehyde
Antigen retrieval step
Enzymatic
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5%

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Verified customer

Submitted Jan 18 2006

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