Anti-UCP3 antibody (ab10985)

Reviews (1)Q&A (2)References (16)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-UCP3 antibody (ab10985)

    Key features and details

    • Rabbit polyclonal to UCP3
    • Suitable for: IHC-P
    • Reacts with: Human
    • Isotype: IgG

    Overview

    • Product name

      Anti-UCP3 antibody
      See all UCP3 primary antibodies

    • Description

      Rabbit polyclonal to UCP3

    • Host species

      Rabbit

    • Tested applications

      Suitable for: IHC-Pmore details

    • Species reactivity

      Reacts with: 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.

      The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

      If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

    Properties

    Applications

    The Abpromise guarantee

    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
    IHC-P
    1/3000.
    Notes
    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

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-UCP3 antibody (ab10985)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-UCP3 antibody (ab10985)
        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 (16)

      Publishing research using ab10985? Please let us know so that we can cite the reference in this datasheet.

      ab10985 has been referenced in 16 publications.

      • Park HJ  et al. The essential role of fructose-1,6-bisphosphatase 2 enzyme in thermal homeostasis upon cold stress. Exp Mol Med 52:485-496 (2020). PubMed: 32203098
      • Lee K  et al. Effect of Dietary Silk Peptide on Obesity, Hyperglycemia, and Skeletal Muscle Regeneration in High-Fat Diet-Fed Mice. Cells 9:N/A (2020). PubMed: 32041272
      • Leger T  et al. EPA is Cardioprotective in Male Rats Subjected to Sepsis, but ALA Is Not Beneficial. Antioxidants (Basel) 9:N/A (2020). PubMed: 32365668
      • Jia X  et al. EGCG Upregulates UCP3 Levels to Protect MIN6 Pancreatic Islet Cells from Interleukin-1ß-Induced Apoptosis. Drug Des Devel Ther 14:4251-4261 (2020). PubMed: 33116413
      • Leger T  et al. Dietary EPA Increases Rat Mortality in Diabetes Mellitus, A Phenomenon Which Is Compensated by Green Tea Extract. Antioxidants (Basel) 8:N/A (2019). PubMed: 31690052
      View all Publications for this product

      Customer reviews and Q&As

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      1-3 of 3 Abreviews or Q&A

      Immunohistochemistry (PFA perfusion fixed frozen sections) abreview for Anti-UCP3 antibody

       Excellent
      Abreviews
      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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      Abcam user community

      Verified customer

      Submitted Jan 18 2006

      Question

      I got your reply last week and I recommend your suggestions to our customer. The customer changed their lysis buffer, blocking time, and denaturing time but unfortunately she could not get a good result. I will attach the data from the customer and you can see the result. There are muscle samples for the positive control but you would not see the band around 34kda. Moreover there is the result of alpha-p38 as a loading control and you can see the correct bands. It means there is no problem with the procedure that the customer did and your product did not work well. So the customer wants to get a credit for it and does not want to spend more time because they are in urgent. I look forward to your reply.

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      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

      BATCH NUMBER 159627 ORDER NUMBER 150 DESCRIPTION OF THE PROBLEM Non-specific band Wrong band size, no band in positive control SAMPLE Rat/ liver, muscle, white adipose tissue PRIMARY ANTIBODY Dilute antibody 1/500 in 3 % skim milk with 0.02 % sodium azide Incubate membrane with antibody at 4 ? for over night. Wash 5 min with TBS?T DETECTION METHOD ECL Plus (Amersham Pharmacia) POSITIVE AND NEGATIVE CONTROLS USED muscle tissue ANTIBODY STORAGE CONDITIONS -20 ? as described in data sheet SAMPLE PREPARATION Lysis buffer ; 50 mM Tris (pH 7.4), 150 mM NaCl, 1 mM EDTA, 25 mM sodium fluoride, 0.2 % Triton X-100, 0.3 % NP 40 , 1 mM sodium orthovanadate, protease inhibitor Lysis with homogenizer and heating samples for 5 min with sample buffer AMOUNT OF PROTEIN LOADED 50 ?, 100 ? ELECTROPHORESIS/GEL CONDITIONS 12 % of SDS PAGE TRANSFER AND BLOCKING CONDITIONS Transfer buffer; 25 mM Tris, 192 mM glycine, 20 % v/v methanol Transfer at 90 V for 50 min and I confirmed protein band with Ponceau. S Blocking agent; 5 % skim milk in TBS?T SECONDARY ANTIBODY Dilute antibody 1/5000 in 3 % skim milk for 1 h. Wash 3 times with TBS?T for 30 min. HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 5 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? I altered dilution factor of antibody 1/1000 to 1/200 ADDITIONAL NOTES I will attach my data. It is urgent experiment in our lab. So, please give me quick response.

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      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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