#27901 XBP-1s (E8C2Z) Mouse mAb
|XBP-1s (E8C2Z) Mouse mAb recognizes endogenous levels of total XBP-1s protein.|
|Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ala285 of human XBP-1s protein.|
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Western blot analysis of extracts from 293T cells, untreated (-) or treated with tunicamycin #12819 (2 μg/ml, 8 hr; +), using XBP-1s (E8C2Z) Mouse mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western blot analysis of extracts from SK-N-AS cells, untreated (-) or treated with thapsigargin #12758 (1 μM, 8 hr; +), using XBP-1s (E8C2Z) Mouse mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Confocal immunofluorescent analysis of SK-N-AS cells, untreated (left) or treated with thapsigargin #12758 (1 μM, 8 hr; right), using XBP-1s (E8C2Z) Mouse mAb (green) and β-Actin (13E5) Rabbit mAb #4970 (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Flow cytometric analysis of 293T cells, untreated (blue) or treated with tunicamycin #12819 (2 μg/ml, 7 hr; green), using XBP-1s (E8C2Z) Mouse mAb (solid lines) or concentration-matched Mouse (E5Y6Q) mAb IgG2a Isotype Control #61656 (dashed lines). Anti-mouse IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4408 was used as a secondary antibody.
Chromatin immunoprecipitations were performed with cross-linked chromatin from 293T cells treated with tunicamycin #12819 (2 μg/ml, 8 hr) and either XBP-1s (E8C2Z) Mouse mAb #27901 or Normal Rabbit IgG #2729 using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human DNAJB9 Exon 1 Primers #79879, human HSPA5 promoter primers, human HNRNPA3 promoter primers, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
Following protein synthesis, secretory, intra-organellar, and transmembrane proteins translocate into the endoplasmic reticulum (ER) where they are post-translationally modified and properly folded. The accumulation of unfolded proteins within the ER triggers an adaptive mechanism known as the unfolded protein response (UPR) that counteracts compromised protein folding (1). The transmembrane serine/threonine kinase IRE1, originally identified in Saccharomyces cerevisiae, is a proximal sensor for the UPR that transmits the unfolded protein signal across the ER membrane (2-4). The human homolog IRE1α was later identified and is ubiquitously expressed in human tissues (5). Upon activation of the unfolded protein response, IRE1α splices X-box binding protein 1 (XBP-1) mRNA through an unconventional mechanism using its endoribonuclease activity (6). This reaction converts XBP-1 from an unspliced XBP-1u isoform to the spliced XBP-1s isoform, which is a potent transcriptional activator that induces expression of many UPR responsive genes (6).
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