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Organelle Detection Western Blot Cocktail (ab133989)

Organelle Detection Western Blot Cocktail (ab133989)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)

Key features and details

  • Assay type: Quantitative
  • Sample type: Adherent cells, Cell culture extracts, Cell Lysate, Nuclear Extracts, Suspension cells, Tissue Extracts, Tissue Homogenate

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Overview

  • Product name

    Organelle Detection Western Blot Cocktail
  • Sample type

    Cell culture extracts, Adherent cells, Suspension cells, Tissue Extracts, Cell Lysate, Tissue Homogenate, Nuclear Extracts
  • Assay type

    Quantitative
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Product overview

    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.

     

  • Tested applications

    Suitable for: WBmore details

Images

  • Western blot - Organelle Detection Western Blot Cocktail (ab133989) - Component Separation
    Western blot - Organelle Detection Western Blot Cocktail (ab133989) - Component Separation

    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

  • Western blot - Organelle Detection Western Blot Cocktail (ab133989) - Cross reactivity
    Western blot - Organelle Detection Western Blot Cocktail (ab133989) - Cross reactivity
    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.
  • Western blot - Organelle Detection Western Blot Cocktail (ab133989) - Cell Fractions
    Western blot - Organelle Detection Western Blot Cocktail (ab133989) - Cell Fractions
    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.

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

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