Total OXPHOS Rodent WB 抗体 Cocktail (ab110413)

製品の概要

  • 製品名Total OXPHOS Rodent WB Antibody Cocktail
  • アッセイタイプQuantitative
  • 種交差性
    交差種: Mouse, Rat, Cow, Human, Cynomolgus Monkey
  • 製品の概要

    ab110413 contains 5 mouse mAbs, one each against CI subunit NDUFB8 (ab110242), CII-30kDa (ab14714), CIII-Core protein 2 (ab14745) CIV subunit I (ab14705) and CV alpha subunit (ab14748) as an optimized premixed cocktail. The kit is suitable for Western Blotting analysis of the relative levels of the 5 OXPHOS complexes in mitochondrial preparations from mouse, rat, human, or bovine sources.

     

    Altered levels of assembly can arise from mutations in individual subunits, mutations in assembly factors for the complex(es), mtDNA depletion or as a result of physiological and or pathological changes e.g. hormone treatment, exercise, diet or oxidative stress.


     

    The mAbs in the cocktail were chosen because they are against a subunit that is labile when its complex is not assembled. Moreover, the combination is readily resolved in SDS-PAGE when the appropriate gel conditions are used (see protocols). Note: Mouse tissue samples can easily be contaminated with antibodies from the animal's blood. To avoid such background bands, use ab110413 in conjunction with an anti-mouse secondary against native antibodies. Also, COXI is a highly hydrophobic protein and appears as a broad band at ~35 kDa (not at its true molecular weight at 57 kDa). It is very sensitive to heating. Therefore, the samples, including the positive control, should not be heated over 50°C before loaded on the gel.

     

    The Western blot cocktail is supplied at a concentration of 1.5 mg/mL.

     

     

  • 特記事項

    Store the antibody cocktail at 4°C and the control sample at -80°C.

  • アプリケーション適用あり: WBmore details

製品の特性

    アプリケーション

    Our Abpromise guarantee covers the use of ab110413 in the following tested applications.

    The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

    アプリケーション Abreviews 特記事項
    WB Use a concentration of 6 µg/ml.

    The antibody cocktail (1.5 mg/mL) should be diluted 250x to a final working concentration of 6.0 µg/mL for Western blotting. Store the antibody cocktail at 4°C and the control sample at -80°C.

    Total OXPHOS Rodent WB Antibody Cocktail 画像

    • Figure 1. Rat liver mitochondria labeled with ab110413 (MS604). The sample in lane 1 was kept at room temperature, whereas the remaining three samples were heated to 37°C, 50°C, and 100°C, respectively. This blot shows that boiling of samples leads to a decrease in signal due to aggregation of proteins therefore heating samples at or close to boiling is not recommended.

    Total OXPHOS Rodent WB Antibody Cocktail (ab110413) 使用論文

    This product has been referenced in:
    • Fiuza-Luces C  et al. Muscle Signaling in Exercise Intolerance: Insights from the McArdle Mouse Model. Med Sci Sports Exerc N/A:N/A (2016). WB . Read more (PubMed: 27031745) »
    • Radde BN  et al. Anacardic Acid, Salicylic Acid, and Oleic Acid Differentially Alter Cellular Bioenergetic Function in Breast Cancer Cells. J Cell Biochem 117:2521-32 (2016). Read more (PubMed: 26990649) »

    See all 94 Publications for this product

    Product Wall

    Abreviews
    Application: WB
    Sample species: Mouse
    1) Sample species: Mouse
    2) Type: Tissue lysate (Gastrocnemius, skeletal muscle)
    3) Amount of protein used: 50ug
    4) Specification: Skeletal Muscle
    5) Reduce/denaturing condition: SDS sample buffer, no boiling. 4-12% gel (Bis/Tris), MOPS
    6) Blocking buffer/concentration/time/temperature:Odyssey Blocking Buffer, Antibody dil. in 4%BSA-TBST/4hrs RT
    7) Antibody dilution: 1:1000
    8) Incubation time/temperature: 4hrs/RT
    9) Diluent: 4% BSA-TBST
    10) Secondary antibody information:LICOR/Goat-anti-mouse/1:20,000
    11) Detection method: LICOR Odyssey
    Username

    Miss. Alice Pollard

    Verified customer

    投稿 Apr 08 2015

    The temperature shown on the WB image is the degrees at which the lysate was denatured prior to loading on the gel (5 min only). We show this as most customers boil the samples before loading. For these antibodies, boiling will actually be counterprodu...

    Read More

    Yes, you can use frozen tissues as sample for all three kits. As a reminder, please follow each individual protocol included with each kit to prepare samples for that particular assay.

    Abreviews
    Complex IV did not appear in skeletal muscle lysates from mouse tissue. Rat heart mitochondrial lysate was used as a positive control (provide from abcam)
    Username

    Abcam user community

    Verified customer

    投稿 Jun 10 2014

    I recommend the following blocking = http://www.abcam.com/10x-blocking-buffer-ab126587.html
    It may be diluted in PBST or TBST to 1X and it may be used for the blocking step prior to primary antibody incubation as well as during primary and second...

    Read More

    Associate Professor

    Excellent Excellent 5/5 (Ease of Use)
    Abreviews
    This product works very well for various mouse tissues, such as liver, islets, brain and heart.
    Username

    Abcam user community

    Verified customer

    投稿 Mar 05 2014

    MitoProfile Cocktail in Cynomolgous Monkey

    Good Excellent 5/5 (Ease of Use)
    Abreviews
    Application: WB
    Sample species: Cynomolgous Monkey
    1) Sample species: Cynomolgous Monkey
    2) Type: Tissue lysate - whole cell
    3) Amount of protein used: 20 ug
    4) Specification: PBMC
    5) Reduce/denaturing condition: Reduced, Denaturing, 12% gel (Bis/Tris)
    6) Blocking buffer/concentration/time/temperature: Milk/5%/18 h/4oC
    7) Antibody dilution: 1:1000
    8) Incubation time/temperature: 18 h/4oC
    9) Diluent: 5% Milk-PBS
    10) Secondary antibody information: Non-Abcam antibody/Goat-anti-mouse/Polyclonal/HRP/1:2000
    11) Detection method: Pierce SuperSignal West Femto
    12) Exposure: 1 s
    13) Observed band size: Specific: 17, 25, 32, 37, 47kDa
    Your feelings/comments on the results: Rating: 4*/Excellent

    *these lanes represent 2 PBMC homogenates, each loaded at 20ug per lane
    Username

    Mr. Daniel Tyrrell

    Verified customer

    投稿 Oct 22 2013

    Abreview for ab110413 (Mouse)

    Excellent Excellent 5/5 (Ease of Use)
    Abreviews
    Application: WB
    Sample species: Mouse

    1. Sample species: Mouse
    2. Type: Tissue lysate - whole
    3. Amount of protein used: 30 ug
    4. Specification: Heart
    5. Reduce/denaturing condition: Reduced, Denaturing, 12% gel
    6. Blocking buffer/concentration/time/temperature: Milk/5%/1 h/25oC
    7. Antibody dilution: 1:1000
    8. Incubation time/temperature: 18 h/4oC
    9. Diluent: Milk-PBS
    10. Secondary antibody information: Non-Abcam antibody/Goat-anti-mouse/Polyclonal/HRP/1:5000
    11. Detection method: ECL+
    12. Exposure: 5 s
    13. Observed band size: Specific: 20, 30, 37, 40, 53 kDa

    Your feelings/comments on the results: Rating: 5*/Excellent
    Username

    Abcam user community

    Verified customer

    投稿 Jun 07 2013

    Abreview for ab110413 (Human)

    Excellent Excellent 5/5 (Ease of Use)
    Abreviews
    Application: WB
    Sample species: Human
    1) Sample species: Human
    2) Type: Tissue lysate - whole
    3) Amount of protein used: 30 ug
    4) Specification: Heart
    5) Reduce/denaturing condition: Reduced, Denaturing, 12% gel
    6) Blocking buffer/concentration/time/temperature: Milk/5%/1 h/25oC
    7) Antibody dilution: 1:1000
    8) Incubation time/temperature: 18 h/4oC
    9) Diluent: Milk-PBS
    10) Secondary antibody information: Non-Abcam antibody/Goat-anti-mouse/Polyclonal/HRP/1:5000
    11) Detection method: ECL+
    12) Exposure: 5 s
    13) Observed band size: Specific: 20, 30, 35, 47, 53 kDa
    Your feelings/comments on the results: Rating: 5*/Excellent
    Username

    Abcam user community

    Verified customer

    投稿 Jun 07 2013

    The aminocaproic acid functions in Bis-tris buffer to aid extraction of membrane-embedded complexes. It is necessary in detecting OXPHOS complexes.

    1-10 of 31 Abreviews or Q&A

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"