PLK1 [p Thr210] Antibody - BSA Free

Western Blot: PLK1 [p Thr210] Antibody [NBP1-78021] - 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.

Immunohistochemistry: PLK1 [p Thr210] Antibody [NBP1-78021] - Used at a 1:200 dilution to detect Plk1 by immunohistochemistry in human breast carcinoma tumor tissue. Tissue was formalin-fixed and paraffin embedded.

Western Blot: PLK1 [p Thr210] Antibody [NBP1-78021] - 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.

Immunohistochemistry: PLK1 [p Thr210] Antibody [NBP1-78021] - Used at a 1:200 dilution to detect Plk1 by immunohistochemistry in human colon 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 breast carcinoma tumor tissue. Tissue was formalin-fixed and paraffin embedded. Personal Communication, Alan Yen, LifeSpanBiosciences, Seattle, WA.

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 aT210 (panel A) and pan reactive apanel 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, aT210 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. Personal communication Hai Jiang, Northwestern Univ.

• Reactivity: Hu, Mu
• Applications: WB, ELISA, IHC
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Format: BSA Free

PLK1 [p Thr210] Antibody - BSA Free Summary

• Description: This antibody is directed against human phosphorylated Plk1 protein. This antibody was affinity purified
  
  Store vial at -20C prior to opening. Aliquot contents and freeze at -20C 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 4C as an undiluted liquid. Dilute only prior to immediate use.
• Immunogen: PLK1 [p Thr210] 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. (Uniprot: P53350)
• Modification: p Thr210
• Marker: Mitosis Marker
• Specificity: Anti-Polo-like Kinase pT210 Antibody is directed against human phosphorylated Plk1 protein. This antibody is specific for phosphorylated human PLK1 protein at the pT210 residue. BLAST analysis indicates 100 % homology of the immunizing sequence with PLK1 homologues from human, chimpanzee, pig, chicken, mouse, rat, Xenopus, dog, mosquito, zebra fish, starfish, sea urchin and fruit fly. Cross reactivity with PLK1 protein homologues from C.elgans and honeybee may also occur as sequence homology varies by one amino acid residue in this sequence. Reactivity with PLK1 protein from other sources is not known. Minimal reactivity is expected with the non-phosphorylated form of the protein.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: PLK1
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • ELISA 1:3000-1:12000
  • Immunohistochemistry 1:200-1:1000
  • Immunohistochemistry-Paraffin 1:200
  • Western Blot 1:200-1:2000
• 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.

Reactivity Notes

BLAST analysis indicates 100 % homology of the immunizing sequence with PLK1 homologues from human, chimpanzee, pig, chicken, mouse, rat, Xenopus, dog, mosquito, zebra fish, starfish, sea urchin and fruit fly. Cross reactivity with PLK1 protein homologues from C.elgans and honeybee may also occur as sequence homology varies by one amino acid residue in this sequence. Reactivity with PLK1 protein from other sources is not known. Minimal reactivity is expected with the non-phosphorylated form of the protein.

Packaging, Storage & Formulations

• Storage: Store at -20C short term. Aliquot and store at -80C long term. Avoid freeze-thaw cycles.
• Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
• Preservative: 0.01% Sodium Azide
• Purity: Immunogen affinity purified

Alternate Names for PLK1 [p Thr210] Antibody - BSA Free

• cell cycle regulated protein kinase
• EC 2.7.11
• EC 2.7.11.21
• PLK
• PLK1
• PLK-1
• polo (Drosophia)-like kinase
• polo like kinase
• polo-like kinase 1polo-like kinase (Drosophila)
• Serine/threonine-protein kinase 13
• serine/threonine-protein kinase PLK1
• STPK13

Background

Polo-like kinase 1, also known as Plk-1, serine-threonine protein kinase 13 and STPK13, may be required for cell division and have a role during G1 or S phase and is associated with the phosphorylation of cyclin B1. Phosphorylation of Plk-1 on T210 contributes to proper mitotic progression. Plk-1 has a nuclear localization and accumulates to a maximum during the G2 and M phases, declines to a nearly undetectable level following mitosis and throughout G1 phase, and then begins to accumulate again during S phase. Plk-1 is a member of the Ser/Thr protein kinase family and the CDC5/Polo subfamily and contains 2 POLO box domains.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Bioinformatics

• Gene Symbol: PLK1
• Entrez:
  • 5347(Human)
• Uniprot:
  • P53350()
  • P53350()
  • P53350()
  • P53350()
  • P53350()