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Immunocytochemistry - Protein phosphatase 1 inhibitor subunit 2 (phospho T72) antibody (ab27850)
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Western blot - Protein phosphatase 1 inhibitor subunit 2 (phospho T72) antibody (ab27850)
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Western blot - Protein phosphatase 1 inhibitor subunit 2 (phospho T72) antibody (ab27850)
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Product Name
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Protein phosphatase 1 inhibitor subunit 2 (phospho T72) antibody
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See all Protein phosphatase 1 inhibitor subunit 2 antibodies (3)...
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Product type
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Primary antibodies
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Description
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Rabbit polyclonal to Protein phosphatase 1 inhibitor subunit 2 (phospho T72)
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Immunogen
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Synthetic phosphopeptide (Human) - derived from the region that contains threonine 72.
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Reacts with
(species key)
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Hu, Ms
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Specificity
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ab27850 recognises Inhibitor-2.
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Tested applications
(see key)
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ICC, WB
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Abreviews
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Application notes
(see key)
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Recommended dilutions
ICC: 1/100.
WB: 1/1000. Predicted molecular weight: 23 kDa.
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
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Positive control
(see definition)
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HeLa or RAW 264.7 cells treated with nocodazole [200ng/ml] for 16 hours.
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Research areas
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Signal Transduction >> Protein Phosphorylation >> Ser / Thr Phosphatases
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Relevance
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Inhibitor-2 (I-2) exists as a heterodimer with protein phosphatase type-1 (PP1), termed MgATP-dependent phosphatase. I-2 binds and forms an inactive complex with PP1 in its unphosphorylated state. This complex is activated through a series of phosphorylations on serine and threonine residues in I-2 that increases phosphatase activity. Multiple kinases have been implicated in phosphorylation of I-2 at threonine 72, namely GSK-3, cdc2 and ERK1. Casein kinase II phosphorylates I-2 at serine residues, which in turn enhances threonine phosphorylation by GSK-3.
Recent evidence has shown that phosphorylation at threonine 72 peaks during prophase of the cell cycle and is localized in the centrosomes. The I-2/PP1 complex also binds neurabin and the kinases Nek2, KPI-2, and Aurora-A. Regulation of I-2/PP1 has been shown to be important in cell cycle, gene expression, ion gating, and neuromodulation.
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Protein phosphatase 1 inhibitor subunit 2 (phospho T72) antibody (ab27850) |
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Database links
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The links below go to external sites and will open in a new browser window |
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Raised in
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Rabbit
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Clonality
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Polyclonal
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Isotype
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IgG
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Purity
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Immunogen affinity purified
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Storage buffer
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Preservative: 0.05% Sodium Azide
Constituents: 50% Glycerol, 1.0 mg/mL BSA (IgG, protease free), Dulbecco's PBS [without magnesium and calcium ions]. pH 7.3 Material safety datasheets (MSDS) for this product:
Glycerol MSDS
Sodium Azide MSDS
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Purification notes
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The antibody was purified from rabbit serum by sequential epitope-specific chromatography. The antibody has been negatively preadsorbed using a non-phosphopeptide corresponding to the site of phosphorylation to remove antibody that is reactive with non-phosphorylated Inhibitor-2. The final product is generated by affinity chromatography using an Inhibitor-2 -derived peptide that is phosphorylated at threonine 72.
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Form
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Liquid
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Storage instructions
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Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C long term. Avoid repeated freeze / thaw cycles.
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At Abcam, we have one centralized database to hold all of our product information, so that everything we know about this Protein phosphatase 1 inhibitor subunit 2 (phospho T72) antibody is on this datasheet. But please do contact us if you would like any reassurance! |
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See below for Protein phosphatase 1 inhibitor subunit 2 (phospho T72) antibody images, references, products related to ab27850 and other tools.
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Protein phosphatase 1 inhibitor subunit 2 (phospho T72) antibody images:
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 Immunocytochemistry - Protein phosphatase 1 inhibitor subunit 2 (phospho T72) antibody (ab27850)
This image shows HeLa cells incubated with ab 27850 at 1/100 dilution with the addition of phosphopeptide immunogen [right], and no peptide [left]. Counterstaining with DAPI and addition of anti rabbit IgG FITC conjugate.
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Western blot - Protein phosphatase 1 inhibitor subunit 2 (phospho T72) antibody (ab27850)
This image shows that only the immunogen phosphopeptide blocks the antibody demonstrating the specificity of the antibody.
All lanes : Protein phosphatase 1 inhibitor subunit 2 (phospho T72) antibody (ab27850) at 1/1000 dilution
Lane 1 : Extracts of RAW 264-7 cells treated with 200ng/ml nocodazole with NO added peptide
Lane 2 : Extracts of RAW 264-7 cells treated with 200ng/ml nocodazole with non-phosphopeptide corresponding to the immunogen
Lane 3 : Extracts of RAW 264-7 cells treated with 200ng/ml nocodazole with generic phosphothreonine containing peptide
Lane 4 : Extracts of RAW 264-7 cells treated with 200ng/ml nocodazole with phosphopeptide immunogen
Secondary
F(ab')2 ant-rabbit IgG HRP conjugate
developed using the ECL technique
Predicted band size : 23 kDa
Observed band size : 44 kDa (why is the actual band size different from the predicted?)
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Western blot - Protein phosphatase 1 inhibitor subunit 2 (phospho T72) antibody (ab27850)
This image shows the induction of IPP2 phosphorlyation at threonine-72 by the addition of nocodazole in both human and mouse systems.
All lanes : Protein phosphatase 1 inhibitor subunit 2 (phospho T72) antibody (ab27850) at 1/1000 dilution
Lane 1 : HeLa cell extract with NO nocodazole
Lane 2 : HeLa cell extract with 200ng/ml nocodazole for 16 hours
Lane 3 : RAW 264.7 cell extract with NO nocodazole
Lane 4 : RAW 264.7 cell extract with 200ng/ml nocodazole for 16 hours
Lane 5 : HeLa cell extract with an antibody recognizing total human IPP2 protein
Lane 6 : HeLa cell extract with an antibody recognizing total human IPP2 protein
Secondary
F(ab')2 ant-rabbit IgG HRP conjugate
developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 23 kDa
Observed band size : 44 kDa (why is the actual band size different from the predicted?)
Additional bands at : 44/46 kDa (possible dimer),46 kDa,75 kDa. We are unsure as to the identity of these extra bands.
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Search PubMed (MEDLINE) for references to Protein phosphatase 1 inhibitor subunit 2
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Protein phosphatase 1 inhibitor subunit 2 (phospho T72) antibody - more information
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