Cell Signaling Technology

Product Pathways - Protein Stability

KLHL12 (2G2) Mouse mAb #9406

Applications Reactivity Sensitivity MW (kDa) Isotype
W H M Mk Endogenous 62 Mouse IgG1

Applications Key:  W=Western Blotting
Reactivity Key:  H=Human  M=Mouse  Mk=Monkey
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

KLHL12 (2G2) Mouse mAb recognizes endogenous levels of total KLHL12 protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a recombinant protein specific to the carboxy terminus of human KLHL12 protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using KLHL12 (2G2) Mouse mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length human KLHL12 (hKLHL12-Myc/DDK; +), using KLHL12 (2G2) Mouse mAb.

Background

Cullins are proteins that function as molecular scaffolds for modular ubiquitin ligases typified by the SCF (Skp1-CUL1-F-box) complex (1-3). The substrate selectivity of these E3 ligases is dictated by a specificity module that binds cullins. In the SCF complex, this module is composed of Skp1, which binds directly to CUL1, and a member of the F-box family of proteins such as Skp2 (1-4). CUL3 has been shown to be required for embryonic development in mammals and Caenorhabditis elegans (5-7) but until recently, its substrate specificity adaptor had yet to be elucidated. It is now recognized that substrate adaptors for CUL3-based ubiquitin ligase complexes contain a conserved BTB/POZ (Pox virus and Zinc finger) domain. This domain, which was initially identified in the Drosophila transcriptional repressors broad complex, tramtrack, and bric-a-brac is present in more than 190 human proteins. BTB proteins contain a variety of putative protein-protein interaction domains, including MATH domains, zinc finger repeats, and kelch repeats (8).There are several lines of evidence suggesting that Kelch-like 12 protein (KLHL12) is a substrate-specific adaptor for the CUL3-based ubiquitin ligase complex. Analysis of the amino acid sequence of KLHL12 reveals an amino-terminal BTB motif, a central linker region, and a carboxy-terminal kelch domain composed of kelch repeats. Furthermore, KLHL12 has been shown to negatively regulate Wnt signaling by binding Disheveled and targeting it for ubiquitin-dependent proteasomal degradation (9). More recently, KLHL12 was shown to drive the assembly of large COPII vesicles by promoting the monoubiquitination of the COPII component Sec31. As a result, CUL3-KLHL12-dependent ubiquitination is essential for collagen export, a step that is required for integrin-dependent mouse embryonic stem cell division (10).

  1. Zheng, N. et al. (2002) Nature 416, 703-9.
  2. Skowyra, D. et al. (1997) Cell 91, 209-19.
  3. Feldman, R.M. et al. (1997) Cell 91, 221-30.
  4. Bai, C. et al. (1996) Cell 86, 263-74.
  5. Singer, J.D. et al. (1999) Genes Dev 13, 2375-87.
  6. Winston, J.T. et al. (1999) Genes Dev 13, 2751-7.
  7. Kurz, T. et al. (2002) Science 295, 1294-8.
  8. Collins, T. et al. (2001) Mol Cell Biol 21, 3609-15.
  9. Angers, S. et al. (2006) Nat Cell Biol 8, 348-57.
  10. Jin, L. et al. (2012) Nature 482, 495-500.

Application References

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!

Companion Products

For Research Use Only. Not For Use In Diagnostic Procedures.

Products