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Anti-PIN1 Antibody EP1479Y
Also for PIN1 (NM_006221)
|A synthetic peptide corresponding to residues near the N-terminus of human Pin1 was used as an immunogen.|
|Human, Mouse, Rat
||0.5~1.0 mg/ml (Lot Dependent)
|WB, IP, FC
||WB: 1:500; ICC: 1:100 - 1:250; FC: 1:50
|PBS 49%,Sodium azide 0.01%,Glycerol 50%,BSA 0.05%|
|Tissue culture supernatant (Protein A or G Sepharose)
|Is unsuitable for IHC-P or IP.
|Homo sapiens peptidylprolyl cis/trans isomerase, NIMA-interacting 1 (PIN1), transcript variant 1|
|Human human rotamase or peptidyl-prolyl cis-trans isomerase Pin1(PPIase) is important in protein folding, assembly and/or transport. Pin1 is nuclear PPIase containing a WW protein interaction domain, and is structurally and functionally related to Ess1/Ptf1, an essential protein in budding yeast. PPIase activity is necessary for Pin1 function in yeast. Depletion of Pin1 from yeast or HeLa cells induces mitotic arrest, whereas HeLa cells overexpressing Pin1 arrest in the G2 phase of the cell cycle. Pin1 is thus an essential PPIase that regulates mitosis presumably by interacting with NIMA and attenuating its mitosis-promoting activity (1). Pin1 is a conserved mitotic regulator essential for the G2/M transition of the eukaryotic cell cycle. Pin1 displays a preference for an acidic residue N-terminal to the isomerized proline bond due to interaction of this acidic side chain with a basic cluster. This raises the possibility of phosphorylation-mediated control of Pin1-substrate interactions in cell cycle regulation (2). |
Wnt Signaling Pathway
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Western blot - Pin1 antibody [EP1479Y]; Anti-Pin1 antibody [EP1479Y] at 1/500 dilution + 293 cell lysate at 10 µg.Secondary.HRP labelled goat anti-rabbit at 1/2000 dilution.Predicted band size : 18 kDa.Observed band size : 18 kDa.
Immunoprecipitation - Anti-Pin1 antibody [EP1479Y]; Pin1 was immunoprecipitated using 0.5mg Hek293 whole cell extract, 10µg of Rabbit monoclonal to Pin1 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-). .The antibody was incubated under agitation with Protein G beads for 10min, Hek293 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab76309.Secondary: Clean blot (HRP conjugate) at 1/1000 dilution.Band: 18kDa: Pin1.
Flow Cytometry-Anti-Pin1 antibody [EP1479Y](ab76309); Overlay histogram showing HeLa cells stained with ab76309 (red line). The cells were fixed with 80% methanol (5 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.