Overview

  • Product name

  • Description

    Mouse polyclonal to UCP2
  • Host species

    Mouse
  • Tested applications

    Suitable for: WBmore details
  • Species reactivity

    Reacts with: Rat, Human
  • Immunogen

    Full length protein, corresponding to amino acids 1-310 of Human UCP2

  • Positive control

    • Human ovary and Rat brain tissue lysate

Properties

Applications

Our Abpromise guarantee covers the use of ab67241 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 1/500 - 1/1000. Detects a band of approximately 33 kDa (predicted molecular weight: 33 kDa).

Target

  • Function

    UCP are mitochondrial transporter proteins that create proton leaks across the inner mitochondrial membrane, thus uncoupling oxidative phosphorylation from ATP synthesis. As a result, energy is dissipated in the form of heat.
  • Tissue specificity

    Widely expressed in adult human tissues, including tissues rich in macrophages. Most expressed in white adipose tissue and skeletal muscle.
  • Sequence similarities

    Belongs to the mitochondrial carrier family.
    Contains 3 Solcar repeats.
  • Cellular localization

    Mitochondrion inner membrane.
  • Information by UniProt
  • Database links

  • Alternative names

    • BMIQ4 antibody
    • Mitochondrial uncoupling protein 2 antibody
    • SLC25A8 antibody
    • Solute carrier family 25 member 8 antibody
    • UCP 2 antibody
    • ucp2 antibody
    • UCP2_HUMAN antibody
    • UCPH antibody
    • Uncoupling protein 2 antibody
    • Uncoupling protein 2 mitochondrial proton carrier antibody
    see all

Images

  • All lanes : Anti-UCP2 antibody (ab67241) at 1/500 dilution

    Lane 1 : UCP2 transfected 293T cell lysate
    Lane 2 : Non transfected 293T cell lysate

    Lysates/proteins at 25 µg/ml per lane.

    Secondary
    All lanes : Goat Anti-Mouse IgG (H&L)-HRP Conjugate at 1/2500 dilution

    Predicted band size: 33 kDa
    Observed band size: 33 kDa

  • Anti-UCP2 antibody (ab67241) at 1/500 dilution + rat brain tissue lysate at 25 µg/ml

    Secondary
    Goat Anti-Mouse IgG (H&L)-HRP Conjugate at 1/2500 dilution

    Predicted band size: 33 kDa
    Observed band size: 33 kDa

References

This product has been referenced in:

  • Chen M  et al. Ursolic acid stimulates UCP2 expression and protects H9c2 cells from hypoxia-reoxygenation injury via p38 signaling. J Biosci 43:857-865 (2018). Read more (PubMed: 30541946) »
  • Chen XF  et al. Effect of puerarin in promoting fatty acid oxidation by increasing mitochondrial oxidative capacity and biogenesis in skeletal muscle in diabetic rats. Nutr Diabetes 8:1 (2018). Read more (PubMed: 29330446) »
See all 18 Publications for this product

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

Application
Western blot
Sample
Ictidomys tridecemlineatus Tissue lysate - whole (Muscle, Brown adipose tissue, Heart, Brain)
Gel Running Conditions
Reduced Denaturing (10%)
Loading amount
30 µg
Specification
Muscle, Brown adipose tissue, Heart, Brain
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 5°C

Amanda Maccannell

Verified customer

Submitted Jul 20 2018

Answer

Thank you for your reply.

Unfortunately due to Canadian custom restrictions we are unable to send you ab77363. I do apologize for this inconvenience but please let me know if there is another antibody that you would like to receive as a free of charge replacement.

I look forward to your reply and helping to resolve this issue.

Free RabMab with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

Read More

Answer

Thank you for your reply.

Regarding the UCP3 results, I agree that the lower band would be the UCP# signal but I am not sure what the higher molecular weight band could be. It may just be non-specific binding of the antibody, or it may be possible that the protein is in a complex. If the protein is in a complex and that is what the higher molecular weight signal is, then I would suggest boiling your samples for 10 minutes to help break these complexes up.

For ab67241, the UCP2 antibody, in one of the western blot images on the datasheet for this product (where rat brain tissue was probed), it does look as though there may be a double band present. Although I believe that as you ran your gel further, you have resolved this double band more than in our testing. I think the best way to determine the results that you are getting is to send a different UCP2 antibody, free of charge, so that you can run the 2 antibodies side by side. The antibody that I was thinking of sending is ab97931:

https://www.abcam.com/UCP2-antibody-ab97931.html



Please let me know how you would you like to continue and if you could also provide either the Abcam order number or the PO# used to purchase the antibody that would be very helpful.

Free RabMab with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

Read More
Application
Western blot
Sample
Rat Tissue lysate - whole (liver)
Loading amount
20 µg
Specification
liver
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%

Abcam user community

Verified customer

Submitted Sep 04 2012

Application
Western blot
Sample
Human Cell lysate - whole cell (HT-29,SW480,Hep2,Hela,HCT116)
Loading amount
30 µg
Specification
HT-29,SW480,Hep2,Hela,HCT116
Gel Running Conditions
Non-reduced Denaturing (12%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Dr. Yan Nan Shen

Verified customer

Submitted Jun 28 2012

Question

LOT NUMBER GR38463-2 ORDER NUMBER 966068 DESCRIPTION OF THE PROBLEM Non-specific band SAMPLE INS-1 cell lysate (rat) PRIMARY ANTIBODY Concentration or dilution : 1:2500, additional notes – 1:1000 Diluent buffer : blocking buffer Incubation time : overnight at 4 degree Incubation temperature: 4 degree Wash Buffer : TTBS Number of washes : 3 times, 5min DETECTION METHOD ECL advance (LAS-3000) POSITIVE AND NEGATIVE CONTROLS USED Positive control : INS-1 cell lyate Negative control : not used ANTIBODY STORAGE CONDITIONS -20C SAMPLE PREPARATION Lysis buffer : 50mM Tri-HCl, pH 7.5, 150 mM NaCl, 1mM EDTA, 0.25% sodium deoxycholate, 1% NP-40 Protease inhibitors: Roche complete protease inhibitor cocktail (cat. No. 11 697 498 001) Phosphatase inhibitors : sigma phosphatase inhibitor cocktail 3 (cat.no. P0044) sigma phosphatase inhibitor cocktail 2 (cat.no. P5726) Boiling for ≥5 min? yes/no : Yes AMOUNT OF PROTEIN LOADED 10ug/lane ELECTROPHORESIS/GEL CONDITIONS 12 % SDS-PAGE TRANSFER AND BLOCKING CONDITIONS Type of membrane : PVDF Protein transfer verified: ponceau stain Blocking agent and concentration : 3% BSA / TTBS Blocking time : 2 hr Blocking temperature : RT SECONDARY ANTIBODY anti-mouse-HRP (santa cruz) Species: goat Isotype: IgG Reacts against: mouse Concentration or dilution : 1:1000 Diluent buffer : blocking buffer Incubation time : 2hr Incubation temperature: RT Fluorochrome or enzyme conjugate: HRP Wash Buffer : TTBS Number of washes : 3 times, 5min HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 3 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? Antibody dilution factor ADDITIONAL NOTES -

Read More
Answer

Thank you and your customer for taking the time to complete our questionnaire and contact us. I am sorry to hear your customer has had difficulty obtaining satisfactory results from this antibody.

The details your customer has kindly provided will enable us to investigate this case for your customer and this is also helpful in our records for monitoring of quality.

Reviewing this case, I would like to offer some suggestions to help optimise the results from ab67241.

1.) UOP2 is located in the membrane of mitochondria. I therefore suggest to use SDS to make sure the protein is included in the lysate and not stuck in the insoluble membrane fraction.

2.)I suggest to decrease the boiling temperature to 65C and to prolong the boiling time up to 10 min. Your target is a multi-pass membrane protein, and these proteins have the unfortunate tendency to aggregate during boiling periods with high temperatures and beta- mercaptoethanol due to their hydrophobic nature.

3.) I also recommend to run a "no primary control" to exclude the possibility that the secondary antibody is producing the background bands

4.) We tested this antibody with rat brain lysates. I suggest to also use this positive control. I could not find any evidence that INS-1 cells show a strong expression of UCP2.

We are happy to offer this technical support. In the event that a product is not functioning in the applications cited on the product data sheet (and the problem has been reported within 6 months of purchase), we will be pleased to provide a credit note, free of charge replacement or refund.

I hope this information is helpful, thank you and your customer for your cooperation. Should the suggestions not improve the results, please do not hesitate to contact me again with the further requested details.

Read More

Answer

Thank you for your inquiry.
Unfortunately, this antibody was not tested for its cross-reactivity to UCP1 and UCP2 experimentally.
We usually assume a cross-reaction when the homology is above 85%. As ab67241 a polyclonal antibody and the full length protein was the immunogen, the antibody might recognize the other family members. But this has not been tested.
I am sorry I could not provide you with definite answer on this occasion and hope this information is nevertheless helpful.

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