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
概述
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产品名称
Organelle Detection Western Blot Cocktail -
样品类型
Cell culture extracts, Adherent cells, Suspension cells, Tissue Extracts, Cell Lysate, Tissue Homogenate, Nuclear Extracts -
检测类型
Quantitative -
种属反应性
与反应: Mouse, Rat, Human -
产品概述
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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经测试应用
适用于: WBmore details
性能
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存放说明
Store at +4°C. Please refer to protocols. -
组件 200 µl Organelle Detection Western Blot Cocktail 1 x 200µl -
研究领域
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细胞定位
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. -
别名
- 38 kDa BFA-dependent ADP-ribosylation substrate
- adenosinetriphosphatase
- aging associated gene 9 protein
see all -
数据库链接
- 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
相关产品
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Compatible Secondaries
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab133989于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
---|---|---|
WB |
Use at an assay dependent concentration.
The antibody cocktail should be diluted 250 X for western blotting. |
说明 |
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WB
Use at an assay dependent concentration. The antibody cocktail should be diluted 250 X for western blotting. |
图片
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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.
实验方案
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.
数据表及文件
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SDS download
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Datasheet download
文献 (2)
ab133989 被引用在 2 文献中.
- Sundaramurthi H et al. Ergolide mediates anti-cancer effects on metastatic uveal melanoma cells and modulates their cellular and extracellular vesicle proteomes. Open Res Eur 3:88 (2023). PubMed: 37981907
- 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