Plk1 (phospho-T210) antibody

• Catalog Number: orb345425
• Category: Antibodies
• Description: Plk1 (phospho-T210) antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ELISA, IHC, WB
• Reactivity: Human, Mouse
• Isotype: IgG
• Immunogen: Anti-Polo-like Kinase pT210 Antibody was produced by repeated immunizations with a synthetic phospho peptide corresponding to an internal region near aa 200-225 of Human Polo-like kinase 1 (Plk1) protein.
• Concentration: 1.0 mg/mL
• Dilution range: ELISA: 1:3,000 - 1:12,000, IHC: 1:200 - 1:1,000, WB: 1:200 - 1:2,000
• Form/Appearance: Liquid (sterile filtered)
• Purity: Anti-Polo-like Kinase pT210 Antibody is directed against human phosphorylated Plk1 protein. This antibody was affinity purified. This antibody is specific for phosphorylated human Plk-1 protein at the pT210 residue. BLAST analysis indicates 100 % homology of the immunizing sequence with Plk-1 homologues from human, chimpanzee, pig, chicken, mouse, rat, Xenopus, dog, mosquito, zebra fish, starfish, sea urchin and fruit fly. Cross reactivity with Plk-1 protein homologues from C.elgans and honeybee may also occur as sequence homology varies by one amino acid residue in this sequence. Reactivity with Plk-1 protein from other sources is not known. Minimal reactivity is expected with the non-phosphorylated form of the protein.
• Conjugation: Unconjugated
• UniProt ID: P53350
• NCBI: 21359873
• Storage: Store vial at -20° C prior to opening. Aliquot contents and freeze at -20° C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4° C as an undiluted liquid. Dilute only prior to immediate use.
• Buffer/Preservatives: 0.01% (w/v) Sodium Azide
• Alternative names: rabbit anti-PLK1 pT210 antibody, Serine/threonine-
• Note: For research use only
• Application notes: This affinity-purified antibody has been tested for use in ELISA, IHC, and by western blot. Specific conditions for reactivity should be optimized by the end user. Expect a band approximately 68 kDa in size corresponding to Plk-1 by western blotting in the appropriate cell lysate or extract. This antibody is positive by IHC on kidney, liver, cancer, thyroid and lymphocyte tissue.
• Expiration Date: 12 months from date of receipt.

Affinity Purified Plk1 pT210 was used at a 1:200 dilution to detect Plk1 by immunohistochemistry in human breast carcinoma tumor tissue. Tissue was formalin-fixed and paraffin embedded.

Affinity Purified Plk1 pT210 was used at a 1:200 dilution to detect Plk1 by immunohistochemistry in human colon carcinoma tumor tissue. Tissue was formalin-fixed and paraffin embedded.

Western blot analysis is shown to detect endogenous and recombinant protein present in HeLa cell lysates transfected with various plk-1 mutation constructs. Blots were reacted with anti-Plk-1 pT210 (panel A) and pan reactive anti-Plk-1 (panel B). Transfected cells were treated with 1 uM nocodazole followed by cell collection, lysate preparation, SDS-PAGE and western blotting. Using a 1:1000 dilution, anti-Plk-1 pT210 detects a single band corresponding to endogenous plk-1, but does not detect recombinant forms of the protein presumably because of a lack of phosphorylation in these mutants.

Western blot analysis is shown using Biorbyt's Affinity Purified anti-Plk-1 pT210 antibody to detect endogenous protein present in a Mouse A20 whole cell lysate (arrowhead). Comparison to a molecular weight marker indicates a band of ~68 kDa corresponding to Plk-1 protein. It is suggested to use a nuclear extract from synchronized cells to greatly increase the abundance of this protein in preparations. The blot was incubated with a 1:500 dilution of the antibody at room temperature followed by detection using standard techniques.