Overview

  • 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.

  • Tested applications

    Suitable for: WBmore details

Properties

Associated products

Applications

Our Abpromise guarantee covers the use of ab139413 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 1/250.

Images

  • Developed 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
  • Developed 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
  • 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

References

This product has been referenced in:

  • Méhul B  et al. Mass spectrometry and DigiWest technology emphasize protein acetylation profile from Quisinostat-treated HuT78 CTCL cell line. J Proteomics 187:126-143 (2018). Read more (PubMed: 30012418) »
See 1 Publication for this product

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