Cell Signaling Technology

Product Pathways - Cytoskeletal Signaling

CIN85 (D1A4) Rabbit mAb #12304

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP H M Mk (R) (Hm) (B) (Dg) (Pg) Endogenous 78, 82 Rabbit IgG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation
Reactivity Key:  H=Human  M=Mouse  R=Rat  Hm=Hamster  Mk=Monkey  B=Bovine  Dg=Dog  Pg=Pig
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

CIN85 (D1A4) Rabbit mAb recognizes endogenous levels of total CIN85 protein. This antibody also detects CIN85 isoform b (CD2BP3) and CIN85 isoform c. This antibody does not cross-react with CMS/CD2AP.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Lys485 of human CIN85 protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using CIN85 (D1A4) Rabbit mAb.

IP

IP

Immunoprecipitation of CIN85 from U-937 cell extracts using either Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or CIN85 (D1A4) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot detection was performed using CIN85 (D1A4) Rabbit mAb.

Isoform Specificity

Isoform Specificity

Western blot analysis of extracts from 293T cells, either mock transfected (-) or transfected with constructs expressing Myc/DDK-tagged full-length human CIN85 isoform a (hCIN85 Isoform a-Myc/DDK; +), CIN85 isoform b (hCIN85 Isoform b-Myc/DDK; +), and CIN85 isoform c (hCIN85 Isoform c-Myc/DDK; +), using CIN85 (D1A4) Rabbit mAb.


Background

CIN85 was independently identified as Cbl-interacting protein of 85 kDa (1), Ruk (regulator of ubiquitous kinase) (2), SETA (SH3 domain-containing gene expressed in tumorigenic astrocytes) (3), and SH3KBP1 (SH3 domain kinase binding protein 1) (4). The genes encoding these proteins were isolated from either human (CIN85), rat (Ruk and SETA), or mouse (SH3KBP1) sources and share between 92% and 97% sequence identity, suggesting that they represent homologues of one gene. Differential promoter usage and alternative splicing is thought to occur in a tissue specific and developmentally regulated manner to generate a complex expression pattern of various transcripts and encoded protein isoforms (5). The main isoform in humans, CIN85, contains three N-terminal SH3 domains, a proline-rich region harboring several P-X-X-P motifs that provide recognition sites for SH3 domain-containing proteins, a PEST sequence implicated in CIN85 degradation, and a C-terminal coiled-coil region for oligomerization (1,2,5,6). The other molecular variants of CIN85 are shorter, N-terminally truncated proteins lacking one, two, or all three of the SH3 domains (1,5,6-8). Proteomic screens suggest that CIN85 is phosphorylated at multiple sites and the role of phosphorylation of some of these sites in regulation of intra- and intermolecular interactions of CIN85 cannot be excluded. CIN85 belongs to the CD2AP/CMS family of adaptor proteins and has been shown to interact with signaling molecules such as c-Cbl, Cbl-b, BLNK, p85/PI3K, GRB2, p130 Cas, and endophilins to coordinate the activity of multiple signaling cascades. Indeed, a growing body of evidence suggests that CIN85 is required for the regulation of a variety of cellular processes including vesicle-mediated transport (9-12), signal transduction (13,14), and cytoskeleton remodelling (15).

  1. Take, H. et al. (2000) Biochem Biophys Res Commun 268, 321-8.
  2. Gout, I. et al. (2000) EMBO J 19, 4015-25.
  3. Borinstein, S.C. et al. (2000) Cell Signal 12, 769-79.
  4. Narita, T. et al. (2001) Cytogenet Cell Genet 93, 133-4.
  5. Buchman, V.L. et al. (2002) Gene 295, 13-17.
  6. Tibaldi, E.V. and Reinherz, E.L. (2003) Int Immunol 15, 313-29.
  7. Bögler, O. et al. (2000) Neuro Oncol 2, 6-15.
  8. Finniss, S. et al. (2004) Biochem Biophys Res Commun 325, 174-82.
  9. Havrylov, S. et al. (2008) Traffic 9, 798-812.
  10. Kowanetz, K. et al. (2004) Mol Biol Cell 15, 3155-66.
  11. Petrelli, A. et al. (2002) Nature 416, 187-90.
  12. Soubeyran, P. et al. (2002) Nature 416, 183-7.
  13. Borthwick, E.B. et al. (2004) J Mol Biol 343, 1135-46.
  14. Peruzzi, G. et al. (2007) J Immunol 179, 2089-96.
  15. Schmidt, M.H. et al. (2003) J Cell Sci 116, 2845-55.

Application References

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

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