Product Pathways - Translational Control
ER Stress Antibody Sampler Kit #9956
|Kit Includes||Quantity||Applications||Reactivity||MW (kDa)||Isotype|
|BiP (C50B12) Rabbit mAb #3177||40 µl||W IHC-P IHC-F||H M||78||Rabbit IgG|
|Calnexin (C5C9) Rabbit mAb #2679||40 µl||W IHC-P IF-IC||H Mk||90||Rabbit|
|Ero1-Lα Antibody #3264||40 µl||W||H||60||Rabbit|
|IRE1α (14C10) Rabbit mAb #3294||40 µl||W IP||H M||130||Rabbit IgG|
|PDI (C81H6) Rabbit mAb #3501||40 µl||W IHC-P IF-IC||H M R Mk||57||Rabbit|
|CHOP (L63F7) Mouse mAb #2895||40 µl||W IP IF-IC||H M R||27||Mouse IgG2a|
|PERK (D11A8) Rabbit mAb #5683||40 µl||W IP IHC-P||H||140||Rabbit IgG|
|Anti-rabbit IgG, HRP-linked Antibody #7074||100 µl||Goat|
|Anti-mouse IgG, HRP-linked Antibody #7076||100 µl||Horse|
Reactivity Key: H=Human M=Mouse R=Rat Mk=Monkey
Species enclosed in parentheses are predicted to react based on 100% sequence homology.
Specificity / Sensitivity
Each antibody in the ER Stress Antibody Sampler Kit detects endogenous levels of its target protein.
Western blot analysis of extracts from PANC1, HepG2 and A204 cells using Calnexin (C5C9) Rabbit mAb #2679.
Western blot analysis of extracts from C6 and A204 cells, untreated or treated with thapsigargin (300 nM) or tunicamycin (24 μg/ml), using CHOP (L63F7) Mouse mAb #2895.
Western blot analysis of extracts from various cell lines using BiP (C50B12) Rabbit mAb #3177.
Western blot analysis of extracts from various cell lines using Ero1-Lα Antibody #3264.
Western blot analysis of extracts from various cell lines using IRE1α (14C10) Rabbit mAb #3294.
Western blot analysis of extracts from various cell types using PDI (C81H6) Rabbit mAb #3501.
The ER Stress Sampler Kit contains reagents to investigate ER stress within the cell. The kit contains enough primary and secondary antibodies to perform four Western blot experiments per primary antibody.
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu156 of human PERK protein, the sequence around Gly584 of human BiP, the sequence around His963 of human IRE1α, the sequence of human PDI and the sequence of human CHOP. Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to a sequence around Ala51 of human calnexin, the sequence around Leu218 of human Ero1-Lα, and the sequence of mouse MBTPS2. Antibodies are purified by protein A and peptide affinity chromatography.
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.During ER stress, the level of CHOP expression is elevated and CHOP functions to mediate programmed cell death (15).
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- Baou, M. et al. (2010) Haematologica 95, 1510-8. Applications: Western Blotting
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* Product-specific protocol.
- 3177 BiP (C50B12) Rabbit mAb
- 3179 Phospho-PERK (Thr980) (16F8) Rabbit mAb
- 2679 Calnexin (C5C9) Rabbit mAb
- 2433 Calnexin Antibody
- 3264 Ero1-Lα Antibody
- 3294 IRE1α (14C10) Rabbit mAb
- 2446 PDI Antibody
- 2895 CHOP (L63F7) Mouse mAb
- 7071 Phototope®-HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
- 7074 Anti-rabbit IgG, HRP-linked Antibody
- 7720 Prestained Protein Marker, Broad Range (Premixed Format)
- 7727 Biotinylated Protein Ladder Detection Pack
- 7003 20X LumiGLO® Reagent and 20X Peroxide
Rabbit Monoclonals Produced Using Epitomics® Technology, U.S. Patent No. 5,675,063.
For Research Use Only. Not For Use In Diagnostic Procedures.