Revision 1

#15100Store at -20C

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.
Applications:

WB, IP

REACTIVITY:

H Mk

SENSITIVITY:

Endogenous

MW (kDa):

64

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q9UJX2

Entrez-Gene Id:

8697

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:100

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Specificity / Sensitivity

APC8 (D5O2D) Rabbit mAb recognizes endogenous levels of total APC8 protein.

Species Reactivity:

Human, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human APC8 protein.

Background

Eukaryotic cell proliferation depends strictly upon the E3 ubiquitin ligase activity of the anaphase promoting complex/cyclosome (APC/C), whose main function is to trigger the transition of the cell cycle from metaphase to anaphase. The APC/C complex promotes the assembly of polyubiquitin chains on substrate proteins in order to target these proteins for degradation by the 26S proteasome (1,2). The vertebrate APC/C complex consists of as many as 15 subunits, including multiple scaffold proteins, two catalytic subunits (APC2, APC11), and a number of proteins responsible for substrate recognition (3). All E3 enzymes, including APC/C, utilize ubiquitin residues activated by E1 enzymes and transferred to E2 enzymes. Research studies indicate that APC/C interacts with the E2 enzymes UBE2S and UBE2C via the RING-finger domain-containing subunit APC11 (4-6). APC/C function relies on multiple cofactors, including an APC/C coactivator formed by the cell division control protein 20 homolog (CDC20) and Cdh1/FZR1. The CDC20/Cdh1 coactivator is responsible for recognition of APC/C substrates through interaction with specific D-box and KEN-box recognition elements within these substrates (7-9).
Anaphase-promoting complex subunit 8 (APC8, CDC23) is a component of the tetratricopeptide repeat (TPR) APC/C sub-complex that also includes APC3 (CDC27) and APC6 (CDC16). APC8 protein associates with APC3 and APC6 to facilitate recruitment of the APC/C coactivation subunits CDC20 and Cdh1/FZR1 (10,11). Research studies suggest that APC8 protein is overexpressed in papillary thyroid cancer and acts as an important regulator of cell cycle progression and cell growth (12).

  1. Qiao, X. et al. (2010) Cell Cycle 9, 3904-12.
  2. Harper, J.W. et al. (2002) Genes Dev 16, 2179-206.
  3. Chang, L. et al. (2014) Nature 513, 388-93.
  4. Carroll, C.W. and Morgan, D.O. (2002) Nat Cell Biol 4, 880-7.
  5. Gmachl, M. et al. (2000) Proc Natl Acad Sci U S A 97, 8973-8.
  6. Leverson, J.D. et al. (2000) Mol Biol Cell 11, 2315-25.
  7. Kraft, C. et al. (2005) Mol Cell 18, 543-53.
  8. Glotzer, M. et al. (1991) Nature 349, 132-8.
  9. Pfleger, C.M. and Kirschner, M.W. (2000) Genes Dev 14, 655-65.
  10. Matyskiela, M.E. and Morgan, D.O. (2009) Mol Cell 34, 68-80.
  11. Thornton, B.R. et al. (2006) Genes Dev 20, 449-60.
  12. Zhang, L. et al. (2011) Endocr Relat Cancer 18, 731-42.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation

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.
XP 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

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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 1
#15100

APC8 (D5O2D) Rabbit mAb

Western Blotting Image 1: APC8 (D5O2D) Rabbit mAb Expand Image
Western blot analysis of extracts from various cell lines using APC8 (D5O2D) Rabbit mAb.
Western Blotting Image 2: APC8 (D5O2D) Rabbit mAb Expand Image
Western blot analysis of extracts from SCC-12 cells expressing either non-targeting shRNA (shNT) or APC8 shRNA (shAPC8), using APC8 (D5O2D) Rabbit mAb (upper) and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Western Blotting Image 3: APC8 (D5O2D) Rabbit mAb Expand Image
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length human APC8 protein (hAPC8-Myc/DDK; +), using APC8 (D5O2D) Rabbit mAb.
Immunoprecipitation Image 1: APC8 (D5O2D) Rabbit mAb Expand Image
Immunoprecipitation of APC8 from MOLT-4 cell extracts. Lane 1 is 10% input, lane 2 is APC8 (D5O2D) Rabbit mAb and lane 3 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900. Western blot analysis was performed using APC8 (D5O2D) Rabbit mAb.