Cell Signaling Technology

Product Pathways - Translational Control

ER Stress Antibody Sampler Kit #9956

Kit Includes Quantity Applications Reactivity MW (kDa) Source
BiP (C50B12) Rabbit mAb # 3177 40 microliters W IHC-P IHC-F H M 78 Rabbit
Phospho-PERK (Thr980) (16F8) Rabbit mAb # 3179 40 microliters W R (M) 170 Rabbit
Calnexin Antibody # 2433 40 microliters W IHC-P IF-IC H 90 Rabbit
Ero1-Lα Antibody # 3264 40 microliters W H 60 Rabbit
IRE1α (14C10) Rabbit mAb # 3294 40 microliters W IP H M R 130 Rabbit
MBTPS2 Antibody # 2157 40 microliters W M (H) (R) 57 Rabbit
PDI Antibody # 2446 40 microliters W IHC-P IF-IC H M R Mk 57 Rabbit
Anti-rabbit IgG, HRP-linked Antibody # 7074 100 Goat

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IHC-F=Immunohistochemistry (Frozen)  IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey

Specificity / Sensitivity

Each antibody in the ER Stress Antibody Sampler Kit detects endogenous levels of its target protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from Neuro2A and PC-12 cells using MBTPS2 Antibody #2157.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Calnexin Antibody #2433.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using PDI Antibody #2446.


Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using BiP (C50B12) Rabbit mAb #3177.

Western Blotting

Western Blotting

Western blot analysis of extracts from AR42J cells, untreated (-) or treated with 1 μM thapsigargin (Tg) for 20 minutes (+), using Phospho-PERK (Thr980) (16F8) Rabbit mAb #3179.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Ero1-Lα Antibody #3264.


Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using IRE1α (14C10) Rabbit mAb #3294.

Source / Purification

Monoclonal antibodies are produced by immunizing rabbits with a synthetic phospho-peptide (KLH-coupled) corresponding to residues surrounding Thr980 of mouse PERK, the sequence around Gly584 of human BiP, and the sequence around His963 of human IRE1α.Polyclonal antibodies are produced by immunizing rabbits with a synthetic peptide (KLH-coupled) derived from a sequence around Ala51 of human calnexin, the sequence around Leu218 of human Ero1-Lα, the sequence around Pro329 of human PDI, and the sequence of mouse MBTPS2. Antibodies are purified by protein A and peptide affinity chromatography.

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 chaperone proteins including calnexin, BiP and protein disulfide isomerase (PDI). Calnexin is an ER membrane, calcium-binding protein that retains newly synthesized glycoproteins inside the ER to ensure proper folding and quality control (1,2). Irregular protein folding within the ER increases BiP synthesis, which binds misfolded proteins to prevent them from forming aggregates and to assist them to refold properly (3).PDI catalyzes the formation and isomerization of disulfide bonds required for a protein to reach its native state (4). Studies have found that the resident ER protein endoplasmic oxidoreductin-1 (Ero1) provides oxidizing potential to the ER in Saccharomyces cerevisiae (5). Ero1-Lα is an ER membrane-associated N-glycoprotein that promotes oxidative protein folding (6). Disruptions of ER homeostasis leads to the accumulation of unfolded proteins. The ER has developed an adaptive mechanism called the unfolded protein response (UPR) to counteract compromised protein folding (7). This is regulated by proteins such as the membrane-bound transcription factor protease site 2 (MBTPS2) and the serine/threonine kinase IRE1 (8-12). The PERK eIF2α kinase is an ER resident transmembrane protein that couples ER stress signals to translation inhibition. ER stress increases PERK activity, which phosphorylates eIF2α to reduce protein translation. PERK activation during ER stress correlates with autophosphorylation of its cytoplasmic kinase domain (13,14). Phosphorylation of PERK at Thr980 can serve as a marker for its activation status.

  1. Bergeron, J.J. et al. (1994) Trends Biochem. Sci. 19, 124-128.
  2. Williams, D.B. (2006) J. Cell Sci. 119, 615-623.
  3. Kohno, K. et al. (1993) Mol. Cell. Biol. 13, 877-890.
  4. Ellgaard, L. and Ruddock, L.W. (2005) EMBO Rep. 6, 28-32.
  5. Frand, A.R. and Kaiser, C.A. (1998) Mol. Cell 1, 161-170.
  6. Cabibbo, A. et al. (2000) J. Biol. Chem. 275, 4827-4833.
  7. Kaufman, R.J. et al. (2002) Nat. Rev. Mol. Cell Biol. 3, 411-421.
  8. Nikawa, J. and Yamashita, S. (1992) Mol. Microbiol. 6, 1441-1446.
  9. Cox, J.S. et al. (1993) Cell 73, 1197-1206.
  10. Mori, K. et al. (1993) Cell 74, 743-756.
  11. Lee, K. et al. (2002) Genes Dev. 16, 452-466.
  12. Shen, J. and Prywes, R. (2004) J. Biol. Chem. 279, 43046-43051.
  13. Harding, H.P. et al. (1999) Nature 397, 271-274.
  14. Shi, Y. et al. (1998) Mol. Cell. Biol. 18, 7499-7509.

Application References

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Companion Products

Rabbit Monoclonals Produced Using Epitomics® Technology, U.S. Patent No. 5,675,063.

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