Journal: Protein science : a publication of the Protein Society

Article Title: Effects of mutation at the D-JH junction on affinity, specificity, and idiotypy of anti-progesterone antibody DB3.

doi: 10.1110/ps.062236806

Figure Lengend Snippet: Figure 2. Structure of the DB3 combining site. (A) Crystal structure of the complex between DB3 and progesterone (Arevalo et al. 1993a) showing some of the contact residues. Light and the heavy chains are in blue and purple, respectively; steroid oxygen atoms in red; steroid A-ring asterisked. (B) Model of site with TrpH100 replaced by arginine. (C) Superposition of A and B.

Article Snippet: Antibody DB3 engineering JOBNAME: PROSCI 15#9 2006 PAGE: 3 OUTPUT: Saturday August 5 02:47:19 2006 csh/PROSCI/118160/ps0622368 1469896x, 2006, 9, D ow nloaded from https://onlinelibrary.w iley.com /doi/10.1110/ps.062236806 by U niversity O f T he Philipines D ilim an M ain L ibrary, W iley O nline L ibrary on [05/04/2024].

Techniques:

Journal: Protein science : a publication of the Protein Society

Article Title: Effects of mutation at the D-JH junction on affinity, specificity, and idiotypy of anti-progesterone antibody DB3.

doi: 10.1110/ps.062236806

Figure Lengend Snippet: Figure 3. Western blot of wild type and mutants of DB3 VH/K. Samples were run on 10% SDS-PAGE and electrotransferred to an Immobilon-P membrane (PVDF); the blot was probed with HRP-coupled sheep anti- mouse k.

Article Snippet: Antibody DB3 engineering JOBNAME: PROSCI 15#9 2006 PAGE: 3 OUTPUT: Saturday August 5 02:47:19 2006 csh/PROSCI/118160/ps0622368 1469896x, 2006, 9, D ow nloaded from https://onlinelibrary.w iley.com /doi/10.1110/ps.062236806 by U niversity O f T he Philipines D ilim an M ain L ibrary, W iley O nline L ibrary on [05/04/2024].

Techniques: Western Blot, SDS Page, Membrane

Journal: Protein science : a publication of the Protein Society

Article Title: Effects of mutation at the D-JH junction on affinity, specificity, and idiotypy of anti-progesterone antibody DB3.

doi: 10.1110/ps.062236806

Figure Lengend Snippet: Figure 4. Antigen-binding and idiotypic properties of periplasmic extracts of E. coli expressing VH/K mutants. (A) Binding activity by ELISA on progesterone-BSA coated wells; (B) Competitive inhibition of binding by free progesterone (100 ng/mL) in ELISA; (C) Competitive inhibition by rabbit polyclonal anti-DB3-idiotype (10 mg/mL). Results are means of duplicates. (d) Deletion; (Co.) negative control.

Article Snippet: Antibody DB3 engineering JOBNAME: PROSCI 15#9 2006 PAGE: 3 OUTPUT: Saturday August 5 02:47:19 2006 csh/PROSCI/118160/ps0622368 1469896x, 2006, 9, D ow nloaded from https://onlinelibrary.w iley.com /doi/10.1110/ps.062236806 by U niversity O f T he Philipines D ilim an M ain L ibrary, W iley O nline L ibrary on [05/04/2024].

Techniques: Binding Assay, Expressing, Activity Assay, Enzyme-linked Immunosorbent Assay, Inhibition, Negative Control

Journal: Protein science : a publication of the Protein Society

Article Title: Effects of mutation at the D-JH junction on affinity, specificity, and idiotypy of anti-progesterone antibody DB3.

doi: 10.1110/ps.062236806

Figure Lengend Snippet: Figure 5. Steroid specificity of DB3 wild-type VH/K and TrpH100Arg mutant. Binding to progesterone-BSA of fixed amounts of wild-type DB3 VH/K (A) and the TrpH100Arg mutant (B) was carried out in the presence of free progesterone, aetiocholanolone, 5a-dihydroprogesterone, 5b-dihydroprogesterone, or testosterone as inhibitors. Results are means of duplicates.

Article Snippet: Antibody DB3 engineering JOBNAME: PROSCI 15#9 2006 PAGE: 3 OUTPUT: Saturday August 5 02:47:19 2006 csh/PROSCI/118160/ps0622368 1469896x, 2006, 9, D ow nloaded from https://onlinelibrary.w iley.com /doi/10.1110/ps.062236806 by U niversity O f T he Philipines D ilim an M ain L ibrary, W iley O nline L ibrary on [05/04/2024].

Techniques: Mutagenesis, Binding Assay

Journal: Protein science : a publication of the Protein Society

Article Title: Effects of mutation at the D-JH junction on affinity, specificity, and idiotypy of anti-progesterone antibody DB3.

doi: 10.1110/ps.062236806

Figure Lengend Snippet: Figure 6. Binding specificity of wild-type DB3 VH/K and the TrpH100Arg mutant for progesterone derivatives. Binding to progester- one-BSA of wild-type DB3 VH/K (A) and the TrpH100Arg mutant (B) was carried out in the presence of free progesterone-3-CMO (C3-CMO), progesterone-6-HMS (C6-HMS), progesterone-11a-HMS (C11a-HMS), or progesterone-21-HMS (C21-HMS). Results are means of duplicates.

Article Snippet: Antibody DB3 engineering JOBNAME: PROSCI 15#9 2006 PAGE: 3 OUTPUT: Saturday August 5 02:47:19 2006 csh/PROSCI/118160/ps0622368 1469896x, 2006, 9, D ow nloaded from https://onlinelibrary.w iley.com /doi/10.1110/ps.062236806 by U niversity O f T he Philipines D ilim an M ain L ibrary, W iley O nline L ibrary on [05/04/2024].

Techniques: Binding Assay, Mutagenesis

Journal: Molecular endocrinology (Baltimore, Md.)

Article Title: Epigenetic regulation of the lncRNA MEG3 and its target c-MET in pancreatic neuroendocrine tumors.

doi: 10.1210/me.2014-1304

Figure Lengend Snippet: Figure 5. Reduced MEG3 and increased c-MET expression in human insulinomas. A, QRT-PCR assay for MEG3 expression using RNA isolated from 23 FFPE sporadic insulinoma specimens and six normal islet preps showing significantly decreased expression of MEG3 in insulinomas (P .005). Gapdh was used the internal control. B and C, Methylation specific PCR assay of DNA isolated from the specimens in panel A showing increased methylation at the CRE site in the human MEG3 promoter in the insulinomas (34% in normal islets increased to 58% in insulinomas). Meth, methylated; unmeth, unmethylated; D, Immunohistochemical detection of insulin and c-MET (representative images of one normal and one tumor) (1 and 2, 400 and 1000 magnification, respectively), showing an insulin-stained normal islet (3 and 4, 400 and 1000 magnification, respectively), showing no staining with the c-MET antibody in the normal islet. Normal islet is outlined with a black dashed line (5 and 6, 1000 magnification), showing positive staining for insulin and c-MET, respectively. Blue is hematoxylin counterstain in the nucleus.

Article Snippet: For immunohistochemistry (IHC) controls, FFPE pancreas sections of normal human pancreas (n 7) were obtained (Abcam, IHC World, Origene, ProSci, US Biomax, Zyagen, and Dr Michael Emmert-Buck of National Cancer Institute).

Techniques: Expressing, Quantitative RT-PCR, Isolation, Control, Methylation, Immunohistochemical staining, Staining