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Anti-PPP1R1A Antibody EP902Y
Also for PPP1R1A (NM_006741)
|A synthetic peptide corresponding to residues in the N-terminus of human IPP-1 was used as an immunogen.|
|Human, Mouse, Rat
||0.5~1.0 mg/ml (Lot Dependent)
|WB, IHC, IF, FC
||WB: 1:100000; IP: 1:50; FC: 1:20; ICC/IF: Use a concentration of 5 ug/ml; IHC-P: Use at an assay dependent dilution
|PBS 49%,Sodium azide 0.01%,Glycerol 50%,BSA 0.05%|
|Tissue culture supernatant (Protein A or G Sepharose)
|Homo sapiens protein phosphatase 1, regulatory (inhibitor) subunit 1A (PPP1R1A)|
|The inhibitor of protein phosphatase 1 (IPP-1, I-1) is an endogenous inhibitor of protein phophastase 1 (PP-1), a signal transducer in many cellular pathways. IPP-1 is a member of heat-stable PP1 inhibitory subunits, which includes DARPP-32, (a homologue of IPP-1) and IPP-2. First discovered in rabbit skeletal muscle, IPP-1 is expressed in various mammalian tissues. IPP-1 inhibition of PP-1 is controlled through phosphorylation at Thr35 by cAMP-dependent protein kinase and a tetra-peptide consensus motif in various PP-1 binding proteins (1). IPP-1 is a hormonal control in muscle contraction, pituitary cell growth, synaptic plasticity by neurotransmitters, and glycogen metabolism (2). In vivo, IPP-1 is phosphorylated at Ser-67 by NCLK, which also initiates inhibition of PP-1 by IPP-1 (3). Triggered by increase in calcium level, IPP-1 is inactivated by PP2B through de-phosphorylation (4). |
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Western blot - Anti-Protein phosphotase inhibitor 1 antibody [EP902Y]; Anti-Protein phosphatase inhibitor 1 [EP902Y] antibody at 1/40000 dilution + Rat brain at 10 µg.Predicted band size : 19 kDa.Observed band size : 27 kDa .
Anti-Protein phosphotase inhibitor 1 antibody [EP902Y]; Immunohistochemical analysis of paraffin-embedded human normal brain tissue using ab40877 diluted 1:100.
Immunocytochemistry/ Immunofluorescence - Anti-Protein phosphotase inhibitor 1 antibody [EP902Y]; ICC/IF image of ab40877 stained PC12 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody overnight at +4°C. The secondary antibody (green) was Alexa Fluor? 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor? 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Flow Cytometry - Anti-Protein phosphotase inhibitor 1 antibody [EP902Y]; Overlay histogram showing SH-SY5Y cells stained with ab40877 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody for 30 min at 22°C. The secondary antibody used was DyLight? 488 goat anti-rabbit IgG (H+L) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) ( 1µg/1x10^6 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in SH-SY5Y cells fixed with 80% methanol/permeabilized in 0.1% PBS-Tween used under the same conditions.