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 applicationsWB more details

Properties

• Storage instructionsStore at +4°C. Please refer to protocols.

• Components	200 µl
  Organelle Detection Western Blot Cocktail	1 x 200µl

• Research Areas
  • Kits/ Lysates/ Other
  • Kits
  • Cell Metabolism Kits
  • Dehydrogenase Kits

  • Kits/ Lysates/ Other
  • Kits
  • Cell Metabolism Kits
  • Other Metabolism Assay

  • Kits/ Lysates/ Other
  • Kits
  • Cell Metabolism Kits
  • Ion and metal assay kits

  • Kits/ Lysates/ Other
  • Kits
  • Epigenetic kits
  • Histone demethylation

Applications

Organelle Detection Western Blot Cocktail images

• 

  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.
• 

  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)
  
  developed using the ECL technique
  
  Performed under reducing conditions.
  
  Exposure time : 5 minutes