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

Key features and details

  • Sample type: Cell culture extracts

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 Fraction Lysate 1 x 200µl
    HeLa Nuclear Fraction Lysate 1 x 200µl

  • Research areas

Images

  • Figure 1: Western blot with Mitochondria Fraction Western Blot Cocktail (ab139416)
    Figure 1: Western blot with Mitochondria Fraction Western Blot Cocktail (ab139416)

    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

  • Figure 2: Western blot with Plasma Membrane Fraction Western Blot Cocktail (ab139413)
    Figure 2: Western blot with Plasma Membrane Fraction Western Blot Cocktail (ab139413)

    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

  • Figure 3. Western blot with Lysosome Fraction Western Blot Cocktail (ab154472)
    Figure 3. Western blot with Lysosome Fraction Western Blot Cocktail (ab154472)

    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

  • Figure 4: Western blot with Organelle Detection Western Blot Cocktail (ab133989)
    Figure 4: Western blot with Organelle Detection Western Blot Cocktail (ab133989)

    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

  • Figure 5: Western blot with ApoTrack™ Cytochrome c Apoptosis WB Antibody Cocktail Western blot with ApoTrack™ Cytochrome c Apoptosis WB Antibody Cocktail
    Figure 5: Western blot with ApoTrack™ Cytochrome c Apoptosis WB Antibody Cocktail Western blot with ApoTrack™ Cytochrome c Apoptosis WB Antibody Cocktail

    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 Cytochrome c Apoptosis Western Blot Antibody 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

     

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