grp94 2104s antibodies  (Cell Signaling Technology Inc)

Journal: Journal of Lipid Research

Article Title: Characterization of a PNLIP variant identified in Amish pediatric patients with congenital pancreatic lipase deficiency

doi: 10.1016/j.jlr.2025.100878

Figure Lengend Snippet: Expression of the PNLIP p.S290N variant induced increased ER stress and the UPR in vitro. HEK 293T cells were transfected to express PNLIP p.S290N and WT. Samples were harvested 68 h post-transfection for immunoblotting and qPCR analysis for ER stress and the UPR markers as described in the Methods. A: Representative immunoblot images for various markers. α-tubulin served as a control. The relative protein abundance for each marker in different cell lysate fractions presented in (A) was quantified relative to WT, with the exception for the insoluble fraction where the values were expressed as relative to S290N. B: Relative BiP protein abundance. C: Relative GRP94 protein abundance. D: Relative mRNA abundance of BiP and CHOP by qPCR. E: A representative image of semi-quantitative PCR analysis of Xbp1 mRNA splicing. F: Quantification of Xbp1 mRNA splicing presented in (E). Mock, empty vector; WT, wild type. The values in quantification graphs are expressed as the mean ± S.D. of 6 or more independent experiments, and One-Way ANOVA Mixed-effects analysis by Tukey's multiple comparisons were performed.

Article Snippet: The following primary antibodies were used: a rabbit polyclonal antibody against human PNLIP generated in our lab (1:5,000) , a rabbit polyclonal antibody against BiP (1:1,000; Cat.: 3183, Cell Signaling), a rabbit polyclonal antibody against GRP94 (1:1,000; Cat.: 2104, Cell Signaling), and a rat monoclonal antibody against α-tubulin (1:2,000; Cat.: sc-53029, Santa Cruz Biotechnology) used as an endogenous control.

Techniques: Expressing, Variant Assay, In Vitro, Transfection, Western Blot, Control, Quantitative Proteomics, Marker, Real-time Polymerase Chain Reaction, Plasmid Preparation