Plasma Membrane Fraction Western Blot Cocktail (ab139413)
Key features and details
- Assay type: Indirect
- Sample type: Cell Lysate, Nuclear Extracts, Tissue Extracts, Tissue Homogenate
Overview
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Product name
Plasma Membrane Fraction Western Blot Cocktail -
Sample type
Tissue Extracts, Cell Lysate, Tissue Homogenate, Nuclear Extracts -
Assay type
Indirect -
Species reactivity
Reacts with: Mouse, Rat, Human -
Product overview
ab139413 contains 3 mouse monoclonal antibodies each targeting a specific organelle marker. The presence of plasma membrane is determined by Anti-Sodium Potassium ATPase 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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Tested applications
Suitable for: WBmore details
Properties
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Storage instructions
Store at +4°C. Please refer to protocols. -
Components 200 µl 250X Plasma Membrane Fraction WB Cocktail 1 x 200µl -
Research areas
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Cellular localization
Sodium Potassium ATPase: Cell membrane. Melanosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV. 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. -
Database links
- Entrez Gene: 2597 Human
- Entrez Gene: 476 Human
- Entrez Gene: 480 Human
- Entrez Gene: 481 Human
- Entrez Gene: 8350 Human
- Entrez Gene: 8351 Human
- Entrez Gene: 8352 Human
- Entrez Gene: 8353 Human
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Images
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Western Blot for Plasma Membrane Antibody Cocktail – Component SeparationDeveloped using the ECL technique; Performed under reducing conditions; Exposure time: 5 mins; All blocking and antibody incubation steps were done in 5% milk, 20 mM Tris-HCl, 0.1% TWEEN-20
Lane 1: Marker
Lanes 2-5: HeLa Whole Cell Lysate – 20 µg
Primary antibodies:
Lane 1: none
Lane 2: Anti- Sodium ATPase antibody – Plasma Membrane Marker
Lane 3: Anti- GAPDH antibody – Cytosolic Membrane Marker
Lane 4: Anti-Histone 3 antibody – Nuclear Membrane Marker
Lane 5: Assembled Plasma Membrane Antibody Cocktail
Secondary: ab131368 at 1/1000 dilution
Predicted Sodium Potassium ATPase band size: 112 kDa
Observed Sodium Potassium ATPase band size: 100 kDa
Predicted GAPDH band size: 37 kDa
Observed GAPDH band size: 37 kDa
Predicted Histone 3 band size: 17 kDa
Observed Histone 3 band size: 17 kD
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Plasma Membrane Western Blot Cocktail Cross ReactivityDeveloped using ECL technique under reducing conditions; exposure time 3 mins; blocking and antibody incubation steps done in 5% milk/TBST
Lanes:
1: Marker
2: Human heart homogenate Tissue Lysate – 20 µg
3: HeLa Cell Lysate – 20 µg
4: Mouse heart homogenate Tissue Lysate - 20 µg
5: NIH3T3 Cell Lysate – 20 µg
6: Rat heart homogenate Tissue Lysate – 20 µg
7: H9C2 cell lysate – 20 µg
All Lanes:
Anti-Sodium ATPase antibody – Plasma Membrane Marker
Anti-GAPDH antibody – Cytosolic Marker
Anti-Histone H3 (di methyl k9) antibody – Nuclear Marker
Secondary: ab131368 at 1/1000 dilution
Predicted Sodium Potassium ATPase band size: 112 kDa
Observed Sodium Potassium ATPase band size: 100 kDa
Predicted GAPDH band size: 37 kDa
Observed GAPDH band size: 37 kDa
Predicted Histone H3 (di methyl k9) band size: 17 kDa
Observed Histone H3 (di methyl k9 band size: 17 kDa
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WB analysis of HeLa cell fractions.Developed using ECL technique, reducing conditions, exposured 5 mins
Blocking and antibody incubations done in 5% milk, 20 mM Tris-HCl, 0.1% TWEEN-20
Lanes:
1: Marker
2: HeLa Whole Cell Lysate - 20 µl
3: HeLa Cytosolic Fraction Lysate - 20 µl
4: HeLa Membrane Fraction Lysate - 20 µl
5: HeLa Nuclear Fraction Lysate - 20 µl
All Lanes:
Anti-Sodium ATPase antibody – Plasma Membrane Marker
Anti-GAPDH antibody – Cytosolic Membrane Marker
Anti-Histone 3 antibody – Nuclear Membrane Marker
Secondary:
Goat polyclonal to Mouse IgG (ab6789) – H&L (HRP)
Predicted Sodium Potassium ATPase band size: 112 kDa
Observed Sodium Potassium ATPase band size: 100 kDa
Predicted GAPDH band size: 37 kDa
Observed GAPDH band size: 37 kDa
Predicted Histone 3 band size: 17 kDa
Observed Histone 3 band size: 17 kDa
