• Assay Type

  Solid Phase Sandwich ELISA

• Format

  96-well strip plate

• Range

  6.25 - 400 pg/mL

• Sufficient Materials

  For five 96-well plates*

• Specificity

  Please see the

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

Ancillary Reagent Kit Available  

DY008, DuoSet ELISA Ancillary Reagent Kit 2 -

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human Ki-67. The suggested diluent is suitable for the analysis of most cell culture supernate, serum, and plasma samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.

Product Features

• Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time

• Development protocols are provided to guide further assay optimization

• Assay can be customized to your specific needs

• Economical alternative to complete kits

Kit Content

• Capture Antibody

• Detection Antibody

• Recombinant Standard

• Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required

DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # ) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.

The components listed above may be purchased separately:

PBS: (Catalog # ), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na₂HPO₄, 1.5 mM KH₂PO₄, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # ), or 0.05% Tween^® 20 in PBS, pH 7.2-7.4

Reagent Diluent: (Catalog # ), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered

Substrate Solution: 1:1 mixture of Color Reagent A (H₂O₂) and Color Reagent B (Tetramethylbenzidine) (Catalog # )

Stop Solution: 2 N H₂SO₄ (Catalog # )

Microplates: R&D Systems (Catalog # )

Plate Sealers: ELISA Plate Sealers (Catalog # )

Preparation and Storage

• Storage

  Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: Ki-67/MKI67

Ki-67 (also MKI67and TSG126) is a 350-370 kDa nuclear protein that belongs to a molecular group comprised of mitotic chromosome-associated proteins. Ki-67 was originally recognized as an antigen associated with the monoclonal Ki-67 antibody raised against Hodgkin's lymphoma nuclear material. Ki-67 is contextually expressed, being potentially found in all cells that are not in the Go phase of the cell cycle. Thus, KI-67 qualifies as a cell proliferation marker. Functionally, Ki-67 is known to interact with 160 kDa Hklp2, a protein that promotes centrosome separation and spindle bipolarity. It also directly interacts with NIFK, and apparently binds to UBF, thus playing a role in rRNA synthesis.

• Entrez Gene IDs:

  4288 (Human); 17345 (Mouse); 291234 (Rat);

• Long Name:

  Antigen Identified by Monoclonal Antibody Ki-67

• Aliases:

  antigen identified by monoclonal Ki-67; antigen KI-67; Ki67; Ki-67; KIA; Marker Of Proliferation Ki-67; MIB-1; MKI67; PPP1R105; proliferation-related Ki-67 antigen; Protein Phosphatase 1; Regulatory Subunit 105; TSG126