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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.
|Organelle Detection Western Blot Cocktail||1 x 200µl|
Our Abpromise guarantee covers the use of ab133989 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use at an assay dependent concentration.
The antibody cocktail should be diluted 250 X for western blotting.
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
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
ab133989 has not yet been referenced specifically in any publications.