Revision 1

#9406Store 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

REACTIVITY:

H M Mk

SENSITIVITY:

Endogenous

MW (kDa):

62

Source/Isotype:

Mouse IgG1

UniProt ID:

#Q53G59

Entrez-Gene Id:

59349

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000

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

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

Species Reactivity:

Human, Mouse, Monkey

Source / Purification

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

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.

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

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.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

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Revision 1
#9406

KLHL12 (2G2) Mouse mAb

Western Blotting Image 1: KLHL12 (2G2) Mouse mAb Expand Image
Western blot analysis of extracts from various cell lines using KLHL12 (2G2) Mouse mAb.
Western Blotting Image 2: KLHL12 (2G2) Mouse 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 KLHL12 (hKLHL12-Myc/DDK; +), using KLHL12 (2G2) Mouse mAb.