MitoSciences (MS1021)

Organelle Detection Western Blot Cocktail (ab133989)

Overview

  • Product nameOrganelle Detection Western Blot Cocktail
  • Sample type
    Cell culture extracts, Adherent cells, Suspension cells, Tissue Extracts, Cell Lysate, Tissue Homogenate, Nuclear Extracts
  • Assay typeQuantitative
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Product overview

    ab133989 (MS1021) 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.

  • Tested applicationsWBmore details

Properties

Applications

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.

Application Abreviews Notes
WB Use at an assay dependent concentration.

The antibody cocktail should be diluted 250 X for western blotting.

Organelle Detection Western Blot Cocktail images

  • 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.

References for Organelle Detection Western Blot Cocktail (ab133989)

ab133989 has not yet been referenced specifically in any publications.

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