HeLa Cell Fractions (Cytoplasmic/Membrane/Nuclear) (ab168542)

Overview

  • Product name
    HeLa Cell Fractions (Cytoplasmic/Membrane/Nuclear)
  • Sample type
    Cell culture extracts
  • Species reactivity
    Reacts with: Human
  • Product overview

    HeLa fractions were prepared using the Cell Fractionation Kit Standard (ab109719)/Membrane Fractionation Kit (ab139409). Using this kit, cells were fractionated into a cytosolic fraction, a membrane (mitochondrial-containing) fraction and a nuclear fraction without any material loss.

  • Notes

    The cytosolic fraction contains cytosolic proteins and is mostly devoid of mitochondrial, ER, plasma membrane, and nuclear proteins. The membrane fraction contains mitochondrial, ER, and plasma membrane proteins, and is mostly devoid of cytosolic and nuclear proteins. The nuclear fraction contains mainly nuclear proteins and is devoid of cytosolic, mitochondrial, ER, and plasma membrane proteins. Each fraction is proportional to the others, allowing the user to analyze equal volumes of each fraction to determine a distribution of a target of interest between these three fractions. The fractions were lysed in SDS sample buffer containing DTT.

  • Tested applications
    Suitable for: WB, SDS-PAGEmore details

Properties

  • Storage instructions
    Store at -80°C. Please refer to protocols.
  • Components 3 units
    HeLa cytosolic fraction lysate 1 x 200µl
    HeLa membrane (mitochondrial) fraction lysate 1 x 200µl
    HeLa nuclear fraction lysate 1 x 200µl
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab168542 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.
SDS-PAGE Use at an assay dependent concentration.

Load 20 µL/Lane

Images

  • Lane 1: MW marker

    Lane 2: HeLa cytosolic fraction

    Lane 3: HeLa membrane

    Lane 4: HeLa nuclear fraction

    All lysates at 20 µL per lane.

    Block and antibody dilution buffer: 5% milk/TBS/0.1% Tween20

    Primary antibodies (all lanes): ab139416 Mitochondria Fraction Western Blot Cocktail (Complex V-a--mitochondrial marker, GAPDH—cytosolic marker, Histone H3 (dimethyl K9)—nuclear marker) 1:250 dilution

     

    Predicted band sizes: 55, 37, 17 kDa

    Observed band sizes: 55, 37, 17 kDa

  • Lane 1: MW marker

    Lane 2: HeLa cytosolic fraction

    Lane 3: HeLa membrane (mitochondrial) fraction

    Lane 4: HeLa nuclear fraction

    All lysates at 20 µL per lane.

    Block and antibody dilution buffer: 5% milk/TBS/0.1% Tween20

    Primary antibodies (all lanes): ab139413 Plasma Membrane Fraction Western Blot Cocktail (Sodium Potassium ATPase—plasma membrane marker, GAPDH—cytosolic marker, Histone H3 (dimethyl K9)—nuclear marker) 1:250 dilution

     

    Predicted band sizes: 112, 37, 17 kDa

    Observed band sizes: 120, 37, 17 kDa

  • Lane 1: MW marker

    Lane 2: HeLa cytosolic fraction

    Lane 3: HeLa membrane (mitochondrial) fraction

    Lane 4: HeLa nuclear fraction

    All lysates at 20 µL per lane.

    Block and antibody dilution buffer: 5% milk/TBS/0.1% Tween20

    Primary antibodies (all lanes): ab154472 Lysosome Fraction Western Blot Cocktail (LAMP2—lysosome marker, GAPDH—cytosolic marker, Histone H3 (dimethyl K9)—nuclear marker) 1:250 dilution

     

    Observed band sizes: 110, 37, 17 kDa

  • Lane 1: MW marker

    Lane 2: HeLa cytosolic fraction

    Lane 3: HeLa membrane (mitochondrial) fraction

    Lane 4: HeLa nuclear fraction

    All lysates at 20 µL per lane.

    Block and antibody dilution buffer: 5% milk/TBS/0.1% Tween20

    Primary antibodies (all lanes): ab133989 Organelle Detection Western Blot Cocktail (Sodium Potassium ATPase—plasma membrane marker, Complex V-a--mitochondrial marker, GAPDH—cytosolic marker, Histone H3 (dimethyl K9)—nuclear marker) 1:250 dilution

     

    Predicted band sizes: 112, 55, 37, 17 kDa

    Observed band sizes: 100, 55, 37, 17 kDa

  • Lane 1: MW marker
    Lane 2: HeLa cytosolic fraction
    Lane 3: HeLa membrane (mitochondrial) fraction
    Lane 4: HeLa nuclear fraction
    All lysates at 2 µL per lane.
    Block and antibody dilution buffer: 5% milk/PBS
    Primary antibodies (all lanes): ab110415 ApoTrack™ Cytochrome c Apoptosis Western Blot Cocktail (Complex V-a--inner mitochondrial membrane marker, PDH E1-a--mitochondiral matrix marker, GAPDH—cytosolic marker, Cytochrome c) 1:250 dilution

    Predicted band sizes: 55, 43, 37, 12 kDa
    Observed band sizes: 55, 43, 37, 12 kDa

     

References

ab168542 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"

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