MLF1 Interacting Protein [p Thr78] Antibody - BSA Free

Western Blot: MLF1 Interacting Protein [p Thr78] Antibody [NBP1-77938] - Shows detection of MLF1IP phosphorylated at Thr78. HeLa cells were co-infected with the indicated adenoviruses expressing GFP-tagged Plk1 or PBIP1. Blots were probed with the anti-MLF1IP pT78 antibody, stripped, and then reprobed with anti-GFP antibody (KA/G & Park, et al., 2006).

Immunohistochemistry: MLF1 Interacting Protein [p Thr78] Antibody [NBP1-77938] - Used at 20 g/ml to detect signal in a variety of tissues including multi-human, multi-brain and multi-cancer slides. This image shows moderately positive staining of mitotic cells in colon adenocarcinoma at 60X. Tissue was formalin-fixed and paraffin embedded. The image shows localization of the antibody as the precipitated red signal, with a hematoxylin purple nuclear counterstain. Personal Communication, Tina Roush, LifeSpanBiosciences, Seattle, WA.

Immunohistochemistry: MLF1 Interacting Protein [p Thr78] Antibody [NBP1-77938] - Analysis of MLF1IP pT78 at the kinetochores of HeLa cells in different phases of the cell cycle. Fluorescent signals were detectable at the kinetochores as early as G2, became most abundant in prophase cells with a discernible nuclear envelope, and gradually diminished as cells proceeded through mitosis (Kang & Park, et al., 2006).

affinity purified anti-MLF1IP pT78 antibody was used at 20 ug/ml to detect signal in a variety of tissues including multi-human, multi-brain and multi-cancer slides. This image shows moderately positive staining of mitotic cells in colon adenocarcinoma at 60X. Tissue was formalin-fixed and paraffin embedded. The image shows localization of the antibody as the precipitated red signal, with a hematoxylin purple nuclear counterstain. Personal Communication, Tina Roush, LifeSpan Biosciences, Seattle, WA.

• Reactivity: Hu
• Applications: WB, DB, ELISA, ICC/IF, IHC
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Format: BSA Free

MLF1 Interacting Protein [p Thr78] Antibody - BSA Free Summary

• Description: This product was affinity purified from monospecific antiserum by immunoaffinity chromatography using phospho-peptide coupled to agarose beads followed by solid phase adsorption against the non-phospho peptide
  
  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: This affinity purified MLF1 Interacting Protein [p Thr78] Antibody was prepared from whole rabbit serum produced by repeated immunizations with a synthetic peptide corresponding to amino acids surrounding Thr78 of human MLF1 Interacting Protein protein. The immunogen peptide is phosphorylated at Thr78. (Uniprot: Q71F23)
• Modification: p Thr78
• Specificity: This antibody is specific for human MLF1 Interacting Protein protein phosphorylated at Thr78.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: CENPU
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Dot Blot
  • ELISA 1:5000-1:25000
  • Immunocytochemistry/ Immunofluorescence 1:10-1:500
  • Immunohistochemistry 20 ug/ml
  • Immunohistochemistry-Paraffin 1:10-1:500
  • Western Blot 1:500-1:2000
• Application Notes: This affinity purified antibody has been tested for use in ELISA, western blotting, IF, and IHC. Specific conditions for reactivity should be optimized by the end user. Expect a band approximately 65 kDa in size corresponding to MLF1IP protein by western blotting in the appropriately stimulated tissue, cell lysate or extract.

Reactivity Notes

A BLAST analysis was used to suggest cross-reactivity with MLF1 Interacting Protein protein from human, dog, bovine and chimpanzee based on 100% homology with the immunizing sequence. Expect partial reactivity with homologues from rat and mouse (90% homology). Reactivity against homologues from other sources is not known.

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 MLF1 Interacting Protein [p Thr78] Antibody - BSA Free

• CENP-50
• CENP-U
• CENPU50
• CENPUMLF1-interacting protein
• centromere protein of 50 kDa
• centromere protein U
• FLJ23468
• ICEN24
• Interphase centromere complex protein 24
• KLIP1CENP50
• KSHV latent nuclear antigen interacting protein 1
• KSHV latent nuclear antigen-interacting protein 1
• MLF1 interacting protein
• PBIP1
• Polo-box-interacting protein 1

Background

Myeloid leukemia factor-1 (MLF1) Interacting Protein (also known as PBIP1, MLF1IP1, KLIP1 or KSHV latent nuclear antigen interacting protein 1) is a novel polo-like kinase 1 (Plk1) substrate. Plk1 phosphorylation of MLF1IP induces ubiquitination and degradation of MLF1IP prior to the metaphase/ anaphase transition. Several Plk1-dependent phosphorylation sites have been identified on MLF1IP by mass spectrometry. Mutations of these sites stabilize MLF1IP and inhibit mitotic progression. Subsequent in vitro and in vivo MLF1IP phosphorylation and stability assays have revealed that phosphorylation of Thr78 is critical for triggering Plk1-dependent MLF1IP degradation. Expression of a non-degradable Thr78Ala mutant was sufficient to induce a mitotic block. Timely phosphorylation of MLF1IP on Thr78 by Plk1 is critical for eliminating the MLF1IP-imposed mitotic block prior to anaphase onset. MLF1IP is speculated to be a novel tumor suppressor,

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.

Publications for MLF1 Interacting Protein Antibody (NBP1-77938)(1)

Publication using NBP1-77938

• Kang YH, Park JE, Yu LR et al. Self-regulated Plk1 recruitment to kinetochores by the Plk1-PBIP1 interaction is critical for proper chromosome segregation. Mol Cell 2006-11-01 [PMID: 17081991]

Bioinformatics

• Gene Symbol: CENPU
• Uniprot:
  • NP_078905(Human)
  • Q71F23(Human)