Cell Signaling Technology
XP Monoclonal Antibody

Product Pathways - Vesicle Trafficking

RCAS1 (D2B6N) XP® Rabbit mAb #12290

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP IF-IC F H M R (Mk) Endogenous 32 Rabbit IgG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  R=Rat  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

RCAS1 (D2B6N) XP® Rabbit mAb recognizes endogenous levels of total RCAS1 protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly147 of human RCAS1 protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using RCAS1 (D2B6N) XP® Rabbit 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 RCAS1 (hRCAS1-Myc/DDK; +), using RCAS1 (D2B6N) XP® Rabbit mAb.

IP

IP

Immunoprecipitation of RCAS1 from HCT 116 extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or RCAS1 (D2B6N) XP® Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using RCAS1 (D2B6N) XP® Rabbit mAb.


Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Jurkat cells using RCAS1 (D2B6N) XP® Rabbit mAb (blue) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (red).

IF-IC

IF-IC

Confocal immunofluorescent analysis of MCF7 cells, untreated (left) or treated with Brefeldin A #9972 (5 μg/ml, 1 hr; right), using RCAS1 (D2B6N) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Background

Receptor binding cancer antigen expressed on SiSo cells (RCAS1) is also known as estrogen receptor-binding fragment-associated gene 9 (EBAG9). Originally identified as an estrogen-inducible gene (1), RCAS1 was recently found to play a novel role in the adaptive immune response by negatively regulating the cytolytic activity of cytotoxic T lymphocytes (CTLs) (2). RCAS1 is conserved in phylogeny and is ubiquitously expressed in most human tissues and cells (3,4). There is evidence that tissue expression of RCAS1 is increased in a variety of malignancies, including cancers of the gastrointestinal tract, liver, lung, breast, ovary, endometrium, and cervix. Research studies have shown that levels of RCAS1 tissue expression are negatively correlated with the prognosis of patients harboring the aforementioned malignancies (4). It is also noteworthy that research studies have detected elevated levels of RCAS1 in the sera of cancer patients (4). Initial studies indicated that RCAS1 was secreted from cancer cells and functioned as a ligand for a putative receptor expressed on NK cells, as well as T and B lymphocytes, inducing their apoptosis, which enabled cancer cells to evade immune surveillance (5,6). Subsequent studies have identified RCAS1 as a type III transmembrane Golgi protein with the ability to regulate vesicle formation, secretion, and protein glycosylation (2,7-9). Indeed, it has been shown that RCAS1 overexpression negatively regulates the cytolytic function of CTLs by negatively regulating protein trafficking from the trans-Golgi to secretory lysosomes (2). Furthermore, RCAS1 overexpression delays vesicle transport from the ER to Golgi and causes components of the ER quality control and glycosylation machinery to mislocalize. As a consequence, RCAS1 induces the deposition of tumor-associated glycan antigens on the cell surface, which are thought to contribute to tumor pathogenesis through the mediation of adhesion, invasion, and metastasis (8,9).

  1. Watanabe, T. et al. (1998) Mol Cell Biol 18, 442-9.
  2. Rüder, C. et al. (2009) J Clin Invest 119, 2184-203.
  3. Tsuchiya, F. et al. (2001) Biochem Biophys Res Commun 284, 2-10.
  4. Giaginis, C. et al. (2009) Histol Histopathol 24, 761-76.
  5. Matsushima, T. et al. (2001) Blood 98, 313-21.
  6. Nakashima, M. et al. (1999) Nat Med 5, 938-42.
  7. Reimer, T.A. et al. (2005) BMC Cancer 5, 47.
  8. Wolf, J. et al. (2010) FASEB J 24, 4000-19.
  9. Engelsberg, A. et al. (2003) J Biol Chem 278, 22998-3007.

Application References

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

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