Organelle Detection Western Blot Cocktail (ab133989) is part of the reagents, controls & accessories range. Abcam offers high-quality biological reagents and tools including antibodies, proteins, assays, cell lines and lysates.
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- Open research Europe 3:882023Ergolide mediates anti-cancer effects on metastatic uveal melanoma cells and modulates their cellular and extracellular vesicle proteomes.Applications:Unspecified applicationReactive species:Unspecified reactive speciesHusvinee Sundaramurthi,Valentina Tonelotto,Kieran Wynne,Fiona O'Connell,Eve O'Reilly,Marcel Costa-Garcia,Csenger Kovácsházi,Agnes Kittel,Simone Marcone,Alfonso Blanco,Eva Pallinger,Szabolcs Hambalkó,Jose Maria Piulats Rodriguez,Péter Ferdinandy,Jacintha O'Sullivan,David Matallanas,Lasse D Jensen,Zoltán Giricz,Breandán N KennedyPubMed 37981907
- Molecular neurobiology 56:7267-72832019Reverting Metabolic Dysfunction in Cortex and Cerebellum of APP/PS1 Mice, a Model for Alzheimer's Disease by Pioglitazone, a Peroxisome Proliferator-Activated Receptor Gamma (PPARγ) Agonist.Applications:Unspecified applicationReactive species:Unspecified reactive speciesKai Lun Chang et. al.PubMed 31016475
Key facts
- Applications
- WB
- Target
- GAPDH di methyl K9
- Reactive species
- Mouse, Rat, Human
- Form
- Liquid
Reactivity data
| Application | Reactivity | Dilution info | Notes |
|---|---|---|---|
Application WB | Reactivity Reacts | Dilution info - | Notes The antibody cocktail should be diluted 250 X for western blotting. |
Target data
Function
Has both glyceraldehyde-3-phosphate dehydrogenase and nitrosylase activities, thereby playing a role in glycolysis and nuclear functions, respectively (PubMed:11724794, PubMed:3170585). Glyceraldehyde-3-phosphate dehydrogenase is a key enzyme in glycolysis that catalyzes the first step of the pathway by converting D-glyceraldehyde 3-phosphate (G3P) into 3-phospho-D-glyceroyl phosphate (PubMed:11724794, PubMed:3170585). Modulates the organization and assembly of the cytoskeleton (By similarity). Facilitates the CHP1-dependent microtubule and membrane associations through its ability to stimulate the binding of CHP1 to microtubules (By similarity). Component of the GAIT (gamma interferon-activated inhibitor of translation) complex which mediates interferon-gamma-induced transcript-selective translation inhibition in inflammation processes (PubMed:23071094). Upon interferon-gamma treatment assembles into the GAIT complex which binds to stem loop-containing GAIT elements in the 3'-UTR of diverse inflammatory mRNAs (such as ceruplasmin) and suppresses their translation (PubMed:23071094). Also plays a role in innate immunity by promoting TNF-induced NF-kappa-B activation and type I interferon production, via interaction with TRAF2 and TRAF3, respectively (PubMed:23332158, PubMed:27387501). Participates in nuclear events including transcription, RNA transport, DNA replication and apoptosis (By similarity). Nuclear functions are probably due to the nitrosylase activity that mediates cysteine S-nitrosylation of nuclear target proteins such as SIRT1, HDAC2 and PRKDC (By similarity).
Alternative names
GAPD, CDABP0047, OK/SW-cl.12, GAPDH, Glyceraldehyde-3-phosphate dehydrogenase, Peptidyl-cysteine S-nitrosylase GAPDH
Recommended products
Organelle Detection Western Blot Cocktail (ab133989) is part of the reagents, controls & accessories range. Abcam offers high-quality biological reagents and tools including antibodies, proteins, assays, cell lines and lysates.
Key facts
- Applications
- WB
- Target
- GAPDH di methyl K9
- Reactive species
- Mouse, Rat, Human
- Form
- Liquid
- Storage buffer
Preservative: 0.02% Sodium azide
Constituents: Proprietary component, 0.877% Sodium chloride, 0.357% HEPES
Storage
- Shipped at conditions
- Blue Ice
- Appropriate short-term storage conditions
- +4°C
- Appropriate long-term storage conditions
- +4°C
Notes
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.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
Sodium Potassium ATPase also known as Na+/K+-ATPase is an essential membrane enzyme responsible for maintaining the electrochemical gradients of sodium and potassium ions across the plasma membrane. This enzyme has a known molecular mass of approximately 112 kDa. It is widely expressed in most animal cells especially in excitable cells such as neurons and muscle cells. Additionally Na+/K+-ATPase plays an important role in organelle function contributing to processes like nutrient uptake and waste removal.
- Biological function summary
The Na+/K+-ATPase enzyme helps regulate cellular volume and influence cell cycle progression. It forms part of a larger protein complex and is vital for maintaining cellular homeostasis. Beyond its mechanical ion-pumping function it is involved in signaling pathways that impact cell growth and differentiation. Its activity is important for neurotransmission and muscle contraction influencing organelle isolation processes. Na+/K+-ATPase also interacts with other proteins like ATPase inhibitory factor 1 which modulates its function.
- Pathways
Na+/K+-ATPase participates actively in ion transport pathways and signaling cascades. For instance it is important in the ATP hydrolysis-driven transport process which relates directly with the maintenance of membrane potential and cellular excitability. It interacts with the Wnt signaling pathway influencing cellular communication and tissue patterning. Proteins such as phospholemman and Src kinase have functional interactions with Na+/K+-ATPase modulating its activity and influence across these pathways.
- Associated diseases and disorders
Na+/K+-ATPase dysregulation associates with conditions such as hypertension and heart failure. Abnormal function of this enzyme can disrupt ion homeostasis leading to elevated blood pressure or cardiac dysfunction. It also links to neurological disorders like schizophrenia where altered Na+/K+-ATPase activity may affect neurotransmission. The enzyme's interaction with proteins like alpha-synuclein and amyloid-beta plays a role in these pathologies further impacting disease progression and symptomatology.
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3 product images
Western blot - Organelle Detection Western Blot Cocktail (ab133989)
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: Anti-mouse IgG for IP (HRP) 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)
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: Anti-mouse IgG for IP (HRP) ab131368 at 1/1000 dilution.
Western blot - Organelle Detection Western Blot Cocktail (ab133989)
HeLa cell lysate was prepared using the Cell Fractionation Kit Cell Fractionation Kit - Standard 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.
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