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.


    We recommend using a high pH CAPS / PVDF transfer protocol when using this antibody for Western blot. Please contact us for more details.

  • 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 at an assay dependent concentration.

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.

We recommend using a high pH CAPS / PVDF transfer protocol when using this antibody for Western blot. Please contact us for more details.

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.

参考文献

This product has been referenced in:
  • Lopez-Gomez C  et al. Deoxycytidine and deoxythymidine treatment for thymidine kinase 2 deficiency. Ann Neurol N/A:N/A (2017). Mouse . Read more (PubMed: 28318037) »
  • Dias TR  et al. Implications of epigallocatechin-3-gallate in cultured human Sertoli cells glycolytic and oxidative profile. Toxicol In Vitro N/A:N/A (2017). WB . Read more (PubMed: 28323106) »

See all 175 Publications for this product

レビューと Q&A

Oxphos Rodent antibody works on Bovine liver

Excellent Excellent 5/5 (Ease of Use)
Abreviews
Bovine liver 20ug of lysates were loaded on to 4-15% gradient gel.
1:4000 dilution of primary antibody used.
Works great!
Username

Julie Kim

Verified customer

投稿 Apr 19 2017

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...

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Cell lysate: mouse cardiomyocytes

Excellent Excellent 5/5 (Ease of Use)
Abreviews
Blocking step: Milk as blocking agent for 1 hour · Concentration: 5 % · Temperature: RT°C

Dilution: 1/100

Incubation time: 13 hours Temperature: 4°C Diluent: 1x TBS with 0.1% Blocking

Secondary antibody
Name: anti-mouse Conjugation: HRP
Dilution:1/200

Detection method: ECL chemi method
Username

Abcam user community

Verified customer

投稿 Nov 01 2017

WB: Tissue lysate, monkey

Excellent Excellent 5/5 (Ease of Use)
Abreviews
Blocking step: Milk as blocking agent for 1 hour · Concentration: 5 % · Temperature: RT°C

Dilution: 1/100

Incubation time: 13 hours Temperature: 4°C Diluent: 1x TBS with 0.1% Blocking

Secondary antibody
Name: anti-mouse Conjugation: HRP
Dilution:1/200

Detection method: ECL chemi method
Username

Abcam user community

Verified customer

投稿 Nov 01 2017

Total OXPHOS Fetal Sheep heart

Excellent Excellent 5/5 (Ease of Use)
Abreviews
The antibody is supposed to pick up 5 bands, for specific proteins within the 5 OXPHOS complexes. Gels were loaded with 50ug of protein extracted from fetal sheep hearts.
The antibody picked up the 5 bands all at the correct molecular weights.
Username

Abcam user community

Verified customer

投稿 Jan 17 2017

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...

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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

1-10 of 35 Abreviews or Q&A

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

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