Revision 6

#18157Store at +4C

1 Kit

(96 assays)

Species Cross Reactivity

H M Mk

UniProt ID:

#P35226

Entrez-Gene Id:

#648

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Color Storage Temp
Bmi1 Mouse mAb Coated Microwells 52797 96 tests +4C
BMi1 Rabbit Detection mAb 73655 1 ea Green (Lyophilized) +4C
Anti-rabbit IgG, HRP-linked Antibody (ELISA Formulated) 13272 1 ea Red (Lyophilized) +4C
Detection Antibody Diluent 13339 11 ml Green +4C
HRP Diluent 13515 11 ml Red +4C
TMB Substrate 7004 11 ml +4C
STOP Solution 7002 11 ml +4C
Sealing Tape 54503 2 ea +4C
ELISA Wash Buffer (20X) 9801 25 ml +4C
ELISA Sample Diluent 11083 25 ml Blue +4C
Cell Lysis Buffer (10X) 9803 15 ml -20C

*The microwell plate is supplied as 12 8-well modules - Each module is designed to break apart for 8 tests.

Description

The PathScan® Total Bmi1 Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of Bmi1 protein. A Bmi1 mouse mAb has been coated onto the microwells. After incubation with cell lysates, the Bmi1 proteins are captured by the coated antibody. Following extensive washing, a Bmi1 rabbit detection mAb is added to detect captured Bmi1 proteins. Anti-rabbit, HRP-linked antibody is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of absorbance for the developed color is proportional to the quantity of Bmi1 protein.

*Antibodies in this kit are custom formulations specific to kit.

Specificity/Sensitivity

PathScan® Total Bmi1 Sandwich ELISA Kit detects endogenous levels of Bmi1 protein in human cells, as shown in Figure 1. The kit sensitivity is shown in Figure 2. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

Background

The polycomb group (PcG) of proteins contributes to the maintenance of cell identity, stem cell self-renewal, cell cycle regulation, and oncogenesis by maintaining the silenced state of genes that promote cell lineage specification, cell death, and cell-cycle arrest (1-4). PcG proteins exist in two complexes that cooperate to maintain long-term gene silencing through epigenetic chromatin modifications. The first complex, EED-EZH2, is recruited to genes by DNA-binding transcription factors and methylates histone H3 on Lys27. This histone methyl-transferase activity requires the Ezh2, Eed, and Suz12 subunits of the complex (5). Histone H3 methylation at Lys27 facilitates the recruitment of the second complex, PRC1, which ubiquitinylates histone H2A on Lys119 (6). Bmi1 is a component of the PRC1 complex, which together with Ring1 strongly enhances the E3 ubiquitin ligase activity of the Ring2 catalytic subunit (7). Bmi1 plays an important role in the regulation of cell proliferation and senescence through repression of the p16 INK4A and p19 ARF genes and is required for maintenance of adult hematopoietic and neural stem cells (3,4,8-10).

  1. Boyer, L.A. et al. (2006) Nature 441, 349-53.
  2. Lee, T.I. et al. (2006) Cell 125, 301-13.
  3. Park, I.K. et al. (2003) Nature 423, 302-5.
  4. Molofsky, A.V. et al. (2003) Nature 425, 962-7.
  5. Cao, R. and Zhang, Y. (2004) Mol Cell 15, 57-67.
  6. Wang, H. et al. (2004) Nature 431, 873-8.
  7. Cao, R. et al. (2005) Mol Cell 20, 845-54.
  8. Molofsky, A.V. et al. (2005) Genes Dev 19, 1432-7.
  9. Jacobs, J.J. et al. (1999) Nature 397, 164-8.
  10. Jacobs, J.J. et al. (1999) Genes Dev 13, 2678-90.

Background References

    Cross-Reactivity Key

    H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    PathScan is a registered trademark of Cell Signaling Technology, Inc.
    All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

    Limited Uses

    Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

    Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

    Revision 6
    #18157

    PathScan® Total Bmi1 Sandwich ELISA Kit

    PathScan® Total Bmi1 Sandwich ELISA Kit: Image 1 Expand Image
    Figure 1. Detection of Bmi1 protein in HeLa, NIH/3T3, and COS-7 cells is examined by using the PathScan® Total Bmi1 Sandwich ELISA Kit. The absorbance readings at 450 nm are shown in the top figure, while the corresponding Western blot using Bmi1 (D42B3) Rabbit mAb #5856 is shown in the bottom figure.
    PathScan® Total Bmi1 Sandwich ELISA Kit: Image 2 Expand Image
    Figure 2. The relationship between protein concentration of lysates from HeLa cells and the absorbance at 450 nm as detected by the PathScan® Total Bmi1 Sandwich ELISA Kit is shown. Unstarved HeLa cells (85% confluence) were harvested and then lysed.