Phospho-PLK1 (Thr210) Antibody (YA161)

Phospho-PLK1 (Thr210) Antibody (YA161) is a Rabbit-derived and non-conjugated IgG monoclonal antibody, targeting to Phospho-PLK1 (Thr210).

PLK1, a serine/threonine-protein kinase, plays a pivotal role in orchestrating various essential functions throughout the M phase of the cell cycle. It regulates centrosome maturation and spindle assembly, facilitates the removal of cohesins from chromosome arms, inactivates anaphase-promoting complex/cyclosome (APC/C) inhibitors, and governs mitotic exit and cytokinesis. Operating by binding and phosphorylating proteins that are already phosphorylated on specific motifs recognized by the POLO box domains, PLK1 phosphorylates an extensive array of substrates, including BORA, BUB1B/BUBR1, CCNB1, CDC25C, CEP55, ECT2, ERCC6L, FBXO5/EMI1, FOXM1, KIF20A/MKLP2, CENPU, NEDD1, NINL, NPM1, NUDC, PKMYT1/MYT1, PRC1, RACGAP1/CYK4, SGO1, STAG2/SA2, TEX14, TOPORS, p73/TP73, TPT1, WEE1, HNRNPU, and others. PLK1's crucial roles include promoting centrosome functions and bipolar spindle assembly through the phosphorylation of KIZ, NEDD1, and NINL. Additionally, it governs mitotic exit and cytokinesis by phosphorylating CEP55, ECT2, KIF20A/MKLP2, CENPU, PRC1, and RACGAP1. PLK1's involvement extends to kinetochore functions, sister chromatid cohesion, and the regulation of various checkpoint proteins, positioning it as a key player in the intricate network governing cell cycle progression.

Free

P53350

Predicted band size: 68 kDa; Observed band size: 68 kDa

Protein A affinity purified.

Nucleus, Chromosome, Cytoplasm, Midbody.

Non-conjugated

Phosphorylated

AB_3102548

Cell Biology

Primary Antibody; Recombinant Rabbit Monoclonal Antibody

Recombinant, Monoclonal

Rabbit

Human

WB: 1:500-1:1000; ICC/IF: 1:10-1:50; FC: 1:10-1:50

• 
  Western blot analysis of extracts from Hela (lane 2(20μg) and Hela (lane 3(20μg), Calyculin A treated (HY-18983, 100ng/mL, 2hrs) as indicated using Phospho-PLK1 (Thr210)(HY-P80474) Rabbit mAb. Proteins were transferred to a PVDF membrane and blocked with 5% non-fat milk in TBST for 2 hour at room temperature. The primary antibody (1/1000) and Loading control antibody (Beta Actin, HY-P80438, 1/10000) was used in 5% non-fat milk in TBST at 4°C overnight. Goat Anti-Mouse/Rabbit IgG-HRP Secondary Antibody (1/10000) was used for 1 hour at room temperature.
• 
  Immunocytochemistry analysis of RAW264.7 cells labeling Phospho-PLK1 (Thr210) with Phospho-PLK1 (Thr210) antibody (HY-P80474) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with Phospho-PLK1 (Thr210) antibody (HY-P80474) at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
• 
  Flow cytometric analysis of 1X10^6 MCF-7 cells labeling Phospho-PLK1 (Thr210) Antibody(HY-P80474, red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Then stained with the primary antibody at 1/50 dilution for an hour at 4℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L (HY-P8002) was used as the secondary antibody at 1/1,000 dilution for 30 minutes at 4℃. Rabbit IgG Isotype Control (HY-P80879, blue) was used as the isotype control, cells without incubation with primary antibody were used as the unlabeled control (black).

WB, ICC/IF, FC

Liquid

Supplied in 1*TBS (pH7.4), 0.05% BSA and 40% Glycerol. Preservative: 0.05% Sodium Azide.

Stored at -20°C for 1 year. Avoid repeated freeze / thaw cycles.

Shipping with blue ice.

IgG

Endogenous

Synthetic phosphopeptide corresponding to residues surrounding Thr210 of Human PLK1.AA range:181-230.