Organelle Detection Western Blot Cocktail (ab133989)
Key features and details
- Assay type: Quantitative
- Sample type: Adherent cells, Cell culture extracts, Cell Lysate, Nuclear Extracts, Suspension cells, Tissue Extracts, Tissue Homogenate
製品の概要
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製品名
Organelle Detection Western Blot Cocktail -
サンプルの種類
Cell culture extracts, Adherent cells, Suspension cells, Tissue Extracts, Cell Lysate, Tissue Homogenate, Nuclear Extracts -
アッセイタイプ
Quantitative -
種交差性
交差種: Mouse, Rat, Human -
製品の概要
ab133989 contains 4 mAbs each targeting a specific organelle marker. The presence of plasma membrane is determined by Anti-Sodium Potassium ATPase antibody; mitochondrion by Anti-ATP5A antibody; cytosol by Anti-GAPDH; and nucleus by Anti-Histone H3 (di methyl K9). This cocktail is suitable for determining the purity of organelle isolates prior to further characterization.
This product is particularly valuable to researchers working in organelle proteomics. Mass spectrometry is frequently used in this field to determine the protein content of targeted organelle isolates. These isolates are obtained using differential centrifugation, density gradient fractionation, biochemical enrichment, or affinity purification. Unfortunately, the various methods of purification available for organelle isolation are imperfect and leave behind contaminants from undesired regions of the cell. These contaminants are inevitable, but being aware of which contaminants are present is crucial for analysis of mass spectrometry results. The high sensitivity and species cross reactivity of the antibodies in this cocktail will quickly and easily reveal impurities caused by imperfect sample preparation.
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アプリケーション
適用あり: WBmore details
製品の特性
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保存方法
Store at +4°C. Please refer to protocols. -
内容 200 µl Organelle Detection Western Blot Cocktail 1 x 200µl -
研究分野
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細胞内局在
Sodium Potassium ATPase: Cell membrane. Melanosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV. ATP5A: Mitochondrion inner membrane. Peripheral membrane protein. GAPDH: Cytoplasm > cytosol. Nucleus. Cytoplasm > perinuclear region. Membrane. Translocates to the nucleus following S-nitrosylation and interaction with SIAH1, which contains a nuclear localization signal (By similarity). Postnuclear and Perinuclear regions. Histone H3: Nucleus. Chromosome. -
別名
- 38 kDa BFA-dependent ADP-ribosylation substrate
- adenosinetriphosphatase
- aging associated gene 9 protein
see all -
参照データベース
- Entrez Gene: 2597 Human
- Entrez Gene: 476 Human
- Entrez Gene: 480 Human
- Entrez Gene: 481 Human
- Entrez Gene: 498 Human
- Entrez Gene: 8350 Human
- Entrez Gene: 8351 Human
- Entrez Gene: 8352 Human
see all
関連製品
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Compatible Secondaries
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Related Products
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab133989の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
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WB |
Use at an assay dependent concentration.
The antibody cocktail should be diluted 250 X for western blotting. |
特記事項 |
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WB
Use at an assay dependent concentration. The antibody cocktail should be diluted 250 X for western blotting. |
画像
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All blocking and antibody incubation steps were done in 5% milk, 20 mM Tris-HCl, 0.1% TWEEN-20.
Lane 2-6 : Mouse heart homogenate Whole Tissue Lysate 10 µg
Primary antibody:
Lane 2 : Anti-Sodium Potassium ATPase antibody – Plasma Membrane Marker
Lane 3 : Anti-ATP5A antibody – Mitochondrial Marker
Lane 4 : Anti-GAPDH antibody – Cytosolic Marker
Lane 5 : Anti-Histone H3 (di methyl K9) antibody – Nuclear Marker
Lane 6 : Assembled Organelle Detection Cocktail
Secondary: ab131368 at 1/1000 dilution.
Predicted Sodium Potassium ATPase band size : 113 kDa
Observed band size : 85 kDa
Predicted ATP5A band size : 60 kDa
Observed ATP5A band size : 52 kDa
Predicted sample band size : 36 kDa
Observed band size : 36 kDa
Predicted sample band size : 15.5 kDa
Observed band size : 17 kDa -
All blocking and antibody incubation steps were done in 5% milk, 20 mM Tris-HCl, 0.1% TWEEN-20.
All lanes :
Anti-Sodium Potassium ATPase antibody – Plasma Membrane Marker
Anti-ATP5A antibody – Mitochondrial Marker
Anti-GAPDH antibody – Cytosolic Marker
Anti-Histone H3 (di methyl K9) antibody – Nuclear Marker
Lane 1 : Marker
Lane 2 : Human heart homogenate Whole Tissue Lysate - 20 µg
Lane 3 : HeLa Whole Cell Lysate - 20 µg
Lane 4 : Mouse heart homogenate Whole Tissue Lysate - 20 µg
Lane 5 : NIH-3T3 Whole Cell Lysate - 20 µg
Lane 6 : Rat heart homogenate Whole Tissue Lysate - 20 µg
Lane 7 : H9C2 Whole Cell Lysate - 20 µg
Secondary: ab131368 at 1/1000 dilution. -
HeLa cell lysate was prepared using the Cell Fractionation Kit ab109719. All blocking and antibody incubation steps were done in 5% milk, 20 mM Tris-HCl, 0.1% TWEEN-20.
All lanes :
Anti-Sodium Potassium ATPase antibody – Plasma Membrane Marker
Anti-ATP5A antibody – Mitochondrial Marker
Anti-GAPDH antibody – Cytosolic Marker
Anti-Histone H3 (di methyl K9) antibody – Nuclear Marker
Lane 1 : Marker
Lane 2 : HeLa Whole Cell Lysate
Lane 3 : HeLa Cytosolic Fraction Lysate
Lane 4 : HeLa Mitochondrial Fraction Lysate
Lane 5 : HeLa Nuclear Fraction Lysate
Secondary
Goat polyclonal to Mouse IgG – H&L – Pre-Adsorbed (HRP) at 1/10000.
プロトコール
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (1)
ab133989 は 1 報の論文で使用されています。
- Chang KL et al. Reverting Metabolic Dysfunction in Cortex and Cerebellum of APP/PS1 Mice, a Model for Alzheimer's Disease by Pioglitazone, a Peroxisome Proliferator-Activated Receptor Gamma (PPAR?) Agonist. Mol Neurobiol N/A:N/A (2019). PubMed: 31016475