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12718
ER and Golgi-Associated Marker Proteins Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

ER and Golgi-Associated Marker Proteins Antibody Sampler Kit #12718

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ER and Golgi-Associated Marker Proteins Antibody Sampler Kit: Image 1

Confocal immunofluorescent analysis of HeLa cells using Calnexin (C5C9) Rabbit mAb (green). Actin filaments have been labeled using DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

ER and Golgi-Associated Marker Proteins Antibody Sampler Kit: Image 2

Flow cytometric analysis of Hep G2 cells using ERp72 (D70D12) XP® Rabbit mAb (blue) compared to a nonspecific negative control antibody (red).

ER and Golgi-Associated Marker Proteins Antibody Sampler Kit: Image 3

Confocal immunofluorescent analysis of NIH/3T3 cells using PDI (C81H6) Rabbit mAb (green) and β-Actin (8H10D10) Mouse mAb #3700 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

ER and Golgi-Associated Marker Proteins Antibody Sampler Kit: Image 4

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).

ER and Golgi-Associated Marker Proteins Antibody Sampler Kit: Image 5

Confocal immunofluorescent analysis of MCF7 cells using Syntaxin 6 (C34B2) Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5™ (fluorescent DNA dye).

ER and Golgi-Associated Marker Proteins Antibody Sampler Kit: Image 6

After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.

ER and Golgi-Associated Marker Proteins Antibody Sampler Kit: Image 7

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Calnexin (C5C9) Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).

ER and Golgi-Associated Marker Proteins Antibody Sampler Kit: Image 8

Confocal immunofluorescent analysis of PANC-1 cells using ERp72 (D70D12) XP® Rabbit mAb (green) and β-Actin (8H10D10) Mouse mAb #3700 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

ER and Golgi-Associated Marker Proteins Antibody Sampler Kit: Image 9

Immunohistochemical analysis of paraffin-embedded human lung carcinoma using PDI (C81H6) Rabbit mAb.

ER and Golgi-Associated Marker Proteins Antibody Sampler Kit: Image 10

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).

ER and Golgi-Associated Marker Proteins Antibody Sampler Kit: Image 11

Western blot analysis of extracts from various cell lines using Syntaxin 6 (C34B2) Rabbit mAb.

ER and Golgi-Associated Marker Proteins Antibody Sampler Kit: Image 12

Western blot analysis of extracts from PANC1, HepG2 and A204 cells using Calnexin (C5C9) Rabbit mAb.

ER and Golgi-Associated Marker Proteins Antibody Sampler Kit: Image 13

Western blot analysis of extracts from various cell lines using ERp72 (D70D12) XP® Rabbit mAb.

ER and Golgi-Associated Marker Proteins Antibody Sampler Kit: Image 14

Immunohistochemical analysis of paraffin-embedded human lymphoma using PDI (C81H6) Rabbit mAb.

ER and Golgi-Associated Marker Proteins Antibody Sampler Kit: Image 15

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.

ER and Golgi-Associated Marker Proteins Antibody Sampler Kit: Image 16

Immunohistochemical analysis of paraffin-embedded mouse spleen using PDI (C81H6) Rabbit mAb.

ER and Golgi-Associated Marker Proteins Antibody Sampler Kit: Image 17

Western blot analysis of extracts from various cell lines using RCAS1 (D2B6N) XP® Rabbit mAb.

ER and Golgi-Associated Marker Proteins Antibody Sampler Kit: Image 18

Western blot analysis of extracts from various cell types using PDI (C81H6) Rabbit mAb.

ER and Golgi-Associated Marker Proteins Antibody Sampler Kit: Image 19

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.

To Purchase # 12718T
Product # Size Price
12718T
1 Kit  (5 x 20 µl) $ 391

Product Description

The ER and Golgi-Associated Marker Proteins Antibody Sampler Kit contains reagents to examine proteins that help regulate protein folding and vesicle trafficking. This kit includes enough antibody to perform two western blot experiments with each primary antibody.

Specificity / Sensitivity

Each antibody will detect endogenous total levels of their target protein.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to the sequence of human calnexin, the sequence of human PDI, the residues surrounding Met279 of human ERp72 protein, the residues surrounding Gly147 of human RCAS1 protein, and the residues surrounding Tyr140 of mouse syntaxin 6 protein.

Background

Secretory and transmembrane proteins are synthesized on polysomes and translocate into the endoplasmic reticulum (ER) where they are often modified by the formation of disulfide bonds, amino-linked glycosylation, and folding. The ER contains a pool of molecular chaperones to help proteins fold properly. Calnexin is a calcium-binding, ER membrane protein that ensures proper protein folding by retaining newly synthesized glycoproteins within the ER l (1-3). The specificity of calnexin for a subset of glycoproteins is defined by a lectin site, which binds an early oligosaccharide intermediate on the folding glycoprotein (3). Many secretory proteins require the formation of intra- or inter-molecular disulfide bonds to reach their native conformation (4). Protein disulfide isomerase (PDI) catalyzes the formation and isomerization of disulfide bonds during oxidative protein folding (5). The ER-protein Ero1 oxidizes PDI through disulfide exchange, which is followed by PDI-catalyzed disulfide bond formation in folding proteins (6). The ER stress protein 72 (ERp72) contains three thioredoxin homology domains and plays a role in the formation and isomerization of disulfide bonds (7,8).

The tumor-associated antigen RCAS1 negatively regulates cytotoxic T lymphocyte (CTL) cytolytic activity, which impacts vesicle formation, secretion, and protein glycosylation (9-12). Overexpression of RCAS1 impairs CTL cytolytic function by negatively regulating trans-Golgi to secretory lysosome protein trafficking, leading to a delay in ER to Golgi vesicle transport and mislocalization of ER quality control and glycosylation proteins. As a result, RCAS1 induces deposition of tumor-associated glycan antigens on the cell surface, which may contribute to tumor pathogenesis through the mediation of adhesion, invasion, and metastasis (13,14). Syntaxin 6 is a ubiquitously expressed S25C family member of the SNARE proteins (15,16) that is localized to the trans-Golgi and within endosomes. It regulates membrane trafficking by partnering with a variety of other SNARE proteins (17-19) and is involved in the regulation of GLUT4 trafficking, neutrophil exocytosis, and granule secretion (20-22).

  1. Rajagopalan, S. et al. (1994) Science 263, 387-390.
  2. Bergeron, J.J. et al. (1994) Trends Biochem. Sci. 19, 124-128.
  3. Williams, D.B. (2006) J. Cell Sci. 119, 615-623.
  4. Huppa, J.B. and Ploegh, H.L. (1998) Cell 92, 145-148.
  5. Ellgaard, L. and Ruddock, L.W. (2005) EMBO Rep. 6, 28-32.
  6. Tu, B.P. and Weissman, J.S. (2004) J. Cell Biol. 164, 341-346.
  7. Mazzarella, RA et al. (1990) J Biol Chem 265(2), 1094-101.
  8. Satoh, M et al. (2005) Cell Stress Chaperones 10(4), 278-84
  9. Rüder, C. et al. (2009) J Clin Invest 119, 2184-203.
  10. Reimer, T.A. et al. (2005) BMC Cancer 5, 47.
  11. Wolf, J. et al. (2010) FASEB J 24, 4000-19.
  12. Engelsberg, A. et al. (2003) J Biol Chem 278, 22998-3007.
  13. Bock, J.B. et al. (2001) Nature 409, 839-41.
  14. Bock, J.B. et al. (1996) J Biol Chem 271, 17961-5.
  15. Wendler, F. and Tooze, S. (2001) Traffic 2, 606-11.
  16. Bock, J.B. et al. (1997) Mol Biol Cell 8, 1261-71.
  17. Mallard, F. et al. (2002) J Cell Biol 156, 653-64.
  18. Perera, H.K. et al. (2003) Mol Biol Cell 14, 2946-58.
  19. Shewan, A.M. et al. (2003) Mol Biol Cell 14, 973-86.
  20. Martín-Martín, B. et al. (2000) Blood 96, 2574-83.
  21. Wendler, F. et al. (2001) Mol Biol Cell 12, 1699-709.
  22. Kuliawat, R. et al. (2004) Mol Biol Cell 15, 1690-701.

Limited Uses

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For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.