rabbit primary anti β adaptin  (Bio-Rad)

Journal: Journal of the Endocrine Society

Article Title: The Glucagon Receptor Is Expressed in the Frontal Cortex and Impaired Signaling Associates With Cognitive Decline

doi: 10.1210/jendso/bvaf056

Figure Lengend Snippet: The GCGR is expressed in several human brain regions. Ten different brain regions were investigated for GCGR protein expression using Western blotting technique with a previously validated GCGR antibody (cat# ab75240, lot# GR2260811-14, RRID: AB_1523687, Abcam). (A-D) Four cropped blots for GCGR expression (55 kDa) are shown with corresponding positive (GCGR-transfected HEK-293 cells) and negative (mock-transfected HEK-293 cells) controls. Uncropped blots are shown in Supplementary Fig. S1 . Control samples were loaded (1 µg protein) on each blot. Brain region names and brain numbers are shown above (A + B) or below (C + D) the corresponding band for each blot. (E) Protein levels of the GCGR in brain tissue regions (n = 4, loaded with 10 µg protein) measured by Western blotting. Levels of the GCGR were normalized to the total protein concentration for the individual blot and subsequently to the positive control to adjust for interblot variation. (F) Immunohistochemistry staining for the GCGR in the frontal cortex. #1 to #4 correspond to brain #1 to #4 in . Data are presented as means ± SEM. Abbreviations: AU, arbitrary units; GCGR, glucagon receptor; HEK, human embryonic kidney; TP, total protein.

Article Snippet: This was followed by 2 washing steps in PBS and subsequent overnight incubation at 4 °C with rabbit anti-human GCGR polyclonal primary antibody (1:200, cat# ab75240, lot# GR2260811-14, RRID: AB_1523687, Abcam) diluted in PBS.

Techniques: Expressing, Western Blot, Transfection, Control, Protein Concentration, Positive Control, Immunohistochemistry, Staining

Journal: Journal of the Endocrine Society

Article Title: The Glucagon Receptor Is Expressed in the Frontal Cortex and Impaired Signaling Associates With Cognitive Decline

doi: 10.1210/jendso/bvaf056

Figure Lengend Snippet: Immunofluorescent staining shows GCGR positivity in neurons of the human frontal cortex. Representative images from 10 µm sections of a 1 to 2 cm 3 tissue block of frontal cortex from 3 human brains (#1-#3) stained with (A) mouse anti-NeuN (staining for neurons) and rabbit anti-GCGR (brain #3), (B) mouse anti-GFAP (staining for astrocytes) and rabbit anti-GCGR (brain #1), or (C) mouse anti-Iba1 (staining for microglia) conjugated to Alexa Fluor 555 and rabbit anti-GCGR (brain #2). All sections were also stained with anti-mouse Alexa488 and anti-rabbit Alexa594 or anti-rabbit Alexa488 as a control for nonspecific binding of the secondary antibody and DAPI (cyan, cell nuclei) (Supplementary Fig. S2) . Examples of positive colocalization are indicated by the arrows and no colocalization by stippled arrows. The dashed square in each panel highlights the region that is magnified in the corresponding inset located in the lower right corner of the panel. An overview of the employed antibodies is presented in Supplementary Table S1 . Scale bars: 30 µm. Abbreviations: DAPI, 4',6-diamidino-2-phenylindole; GCGR, glucagon receptor; GFAP, glial fibrillary acidic protein; Iba1, ionized calcium binding adaptor molecule 1; NeuN, neuronal nuclear antigen.

Article Snippet: This was followed by 2 washing steps in PBS and subsequent overnight incubation at 4 °C with rabbit anti-human GCGR polyclonal primary antibody (1:200, cat# ab75240, lot# GR2260811-14, RRID: AB_1523687, Abcam) diluted in PBS.

Techniques: Staining, Blocking Assay, Control, Binding Assay

Journal: Journal of the Endocrine Society

Article Title: The Glucagon Receptor Is Expressed in the Frontal Cortex and Impaired Signaling Associates With Cognitive Decline

doi: 10.1210/jendso/bvaf056

Figure Lengend Snippet: Fluorescence in situ hybridization histochemistry demonstrates GCGR mRNA in neurons of the frontal cortex. Representative images from 10 µm sections of a 1 to 2 cm 3 tissue block of frontal cortex from human brains demonstrate GCGR mRNA (green) in A and C using anti-sense probes labeled with digoxigenin and visualized by Alex488-tymamide. Single arrows indicate neurons without lipofuscin (autofluorescence); arrowheads indicate neurons expressing the GCGR mRNA and containing lipofuscin granula. Double arrows indicate GCGR mRNA-negative cells containing lipofuscin granula. Nuclei are counterstained by DAPI (blue). (B) and (D) show the sense-hybridization control sections displaying only lipofuscin granula-containing cells. Scale bars: 15 µm. Abbreviation: GCGR, glucagon receptor.

Article Snippet: This was followed by 2 washing steps in PBS and subsequent overnight incubation at 4 °C with rabbit anti-human GCGR polyclonal primary antibody (1:200, cat# ab75240, lot# GR2260811-14, RRID: AB_1523687, Abcam) diluted in PBS.

Techniques: Fluorescence, In Situ Hybridization, Blocking Assay, Labeling, Expressing, Hybridization, Control

Journal: Journal of the Endocrine Society

Article Title: The Glucagon Receptor Is Expressed in the Frontal Cortex and Impaired Signaling Associates With Cognitive Decline

doi: 10.1210/jendso/bvaf056

Figure Lengend Snippet: Descriptive data for individuals with a GCGR variant (cAMP LoF, G40S, or frameshift) and nonvariant carrier controls at study inclusion (baseline)

Article Snippet: This was followed by 2 washing steps in PBS and subsequent overnight incubation at 4 °C with rabbit anti-human GCGR polyclonal primary antibody (1:200, cat# ab75240, lot# GR2260811-14, RRID: AB_1523687, Abcam) diluted in PBS.

Techniques: Variant Assay

Journal: Journal of the Endocrine Society

Article Title: The Glucagon Receptor Is Expressed in the Frontal Cortex and Impaired Signaling Associates With Cognitive Decline

doi: 10.1210/jendso/bvaf056

Figure Lengend Snippet: Cognitive tests (at baseline) between individuals with a GCGR variant (cAMP LoF, G40S, or frameshift variant) and age- and sex-matched nonvariant carrier controls

Article Snippet: This was followed by 2 washing steps in PBS and subsequent overnight incubation at 4 °C with rabbit anti-human GCGR polyclonal primary antibody (1:200, cat# ab75240, lot# GR2260811-14, RRID: AB_1523687, Abcam) diluted in PBS.

Techniques: Variant Assay

Journal: Oncology Letters

Article Title: Robot‑assisted laparoscopic partial cystectomy for urachal carcinoma: A case report

doi: 10.3892/ol.2025.15003

Figure Lengend Snippet: Representative images of immunohistochemistry results. (A) H&E staining (magnification, ×10; scale bar, 300 µm). (B) H&E staining (magnification, ×20; scale bar, 200 µm). (C) Positive expression of CK20 (magnification, ×20; scale bar, 200 µm). (D) Positive expression of caudal type homeobox 2 (magnification, ×20; scale bar, 200 µm). (E) Ki-67 staining demonstrates ~50% positive cells in the hot-spot area (magnification, ×20; scale bar, 200 µm). (F) Positive membrane-bound expression of β-catenin (magnification, ×20; scale bar, 200 µm). H&E, hematoxylin and eosin.

Article Snippet: Subsequently, sections were incubated overnight at 4°C with rabbit primary antibodies against CK20 (cat. no. ab64090; 1:100; Abcam), caudal type homeobox 2 (CDX2; cat. no. ab101532; 1:100; Abcam), β-catenin (cat. no. ab32572; 1:500; Abcam) and Ki-67 (cat. no. ab15580; 1:100; Abcam).

Techniques: Immunohistochemistry, Staining, Expressing, Membrane

Journal: bioRxiv

Article Title: Defining the chromatin-associated protein landscapes on Trypanosoma brucei repetitive elements using synthetic TALE proteins

doi: 10.1101/2025.04.22.649942

Figure Lengend Snippet: Protein extracted from 427 cell and 427 cells with constructs designed to express 177R-TALE, 70R-TALE, 147R-TALE, ingiR-TALE, NonR-TALE and TelR-TALE fused to Ty and YFP tags integrated at the β-tubulin locus was subject to western analysis using either: A. monoclonal mouse anti-GFP (anti-YFP) or B. anti-BB2 (anti-Ty). The TelR-TALE protein is smaller than the 177R-TALE, 70R-TALE, 147R-TALE, ingiR-TALE and NonR-TALE proteins.

Article Snippet: Rabbit anti-GFP primary antibody (Thermo Fisher Scientific A-11122) was used at 1:500 dilution, and secondary Alexa fluor 568 goat antirabbit antibody (Thermo Fisher Scientific A-11036) was used at 1:1000 dilution.

Techniques: Construct, Western Blot

Journal: bioRxiv

Article Title: Defining the chromatin-associated protein landscapes on Trypanosoma brucei repetitive elements using synthetic TALE proteins

doi: 10.1101/2025.04.22.649942

Figure Lengend Snippet: Bloodstream form Lister 427 T. brucei cells expressing the indicated TALE-YFP fusion proteins fixed and TALE-YFP protein localization detected with anti-GFP primary antibody and Alexa Fluor 568-labeled secondary antibody (red). Nuclear and kinetoplastid (mitochondrial) DNA were stained with DAPI (green). Controls cell expressing telomeric YFP-TRF, centromeric YFP-KKT2 kinetochore protein or wild type Lister 427 cells expressing no YFP, are also shown. Scale bar, 10 μm.

Article Snippet: Rabbit anti-GFP primary antibody (Thermo Fisher Scientific A-11122) was used at 1:500 dilution, and secondary Alexa fluor 568 goat antirabbit antibody (Thermo Fisher Scientific A-11036) was used at 1:1000 dilution.

Techniques: Expressing, Labeling, Staining

Journal: bioRxiv

Article Title: Defining the chromatin-associated protein landscapes on Trypanosoma brucei repetitive elements using synthetic TALE proteins

doi: 10.1101/2025.04.22.649942

Figure Lengend Snippet: Analysis of anti-GFP ChIP-seq data for 147R-TALE, 177R-TALE, 70R-TALE, TelR-TALE and ingiR-TALE, demonstrates that each synthetic protein is enriched on the repeat elements they were designed to recognize, namely CIR147 repeats, Ingi retrotransposons, 177bp repeats, 70bp repeats and telomeric (TTAGGG)n repeats. Enrichments obtained for the YFP-KKT2 kinetochore protein, the TRF telomere repeat binding protein and with a No-Tag control are shown for comparison.

Article Snippet: Rabbit anti-GFP primary antibody (Thermo Fisher Scientific A-11122) was used at 1:500 dilution, and secondary Alexa fluor 568 goat antirabbit antibody (Thermo Fisher Scientific A-11036) was used at 1:1000 dilution.

Techniques: ChIP-sequencing, Binding Assay, Control, Comparison

Journal: bioRxiv

Article Title: Defining the chromatin-associated protein landscapes on Trypanosoma brucei repetitive elements using synthetic TALE proteins

doi: 10.1101/2025.04.22.649942

Figure Lengend Snippet: A. Telomeric repeat (TTAGGG) n sequence (top) and 70bp repeat consensus sequence (bottom). Sequences that TelR-TALE and 70R-TALE were designed to bind is indicated. Deletion of TelR-TALE recognition modules following integration in T. bruce i results in recognition of AGGGTTAG rather than the full 15 bp target equence. B. Anti-GFP ChIP-seq for cells expressing TelR-TALE-YFP, YFP-TRF or 70R-TALE–YFP proteins, or 427 cells expressing no YFP tagged protein. ANti-GFP ChIP-seq enrichment profiles are shown for telomeric blood stream expression sites (BES) BES1 (top) and BES5 (bottom). Diagrams show the position of telomeric (TTAGGG) n (black chevrons), VSG genes (blue) and upstream 70 bp repeats (green bars).

Article Snippet: Rabbit anti-GFP primary antibody (Thermo Fisher Scientific A-11122) was used at 1:500 dilution, and secondary Alexa fluor 568 goat antirabbit antibody (Thermo Fisher Scientific A-11036) was used at 1:1000 dilution.

Techniques: Sequencing, ChIP-sequencing, Expressing

Journal: bioRxiv

Article Title: Defining the chromatin-associated protein landscapes on Trypanosoma brucei repetitive elements using synthetic TALE proteins

doi: 10.1101/2025.04.22.649942

Figure Lengend Snippet: A. CIR147 repeat consensus sequence. Sequence that 147R-TALE-YFP was designed to bind is indicated. B. Comparison of sequences enriched in YFP-KKT2 (purple) and 147R-TALE-YFP (blue) anti-GFP ChIP-seq for chromosomes 1, 3, 4, 5 and 8. DNA from all centromeres are enriched in YFP-KKT2 anti-GFP ChIP-seq whereas only CIR147 repeats at centomeres on chromosomes 4, 5 and 8 are enriched in 147R-TALE-YFP anti-GFP ChIP-seq. C. Violin plot demonstrating the relative enrichment of YFP-KKT2 (purple) and 147R-TALE-YFP (blue) over all main chromosome centromere regions.

Article Snippet: Rabbit anti-GFP primary antibody (Thermo Fisher Scientific A-11122) was used at 1:500 dilution, and secondary Alexa fluor 568 goat antirabbit antibody (Thermo Fisher Scientific A-11036) was used at 1:1000 dilution.

Techniques: Sequencing, Comparison, ChIP-sequencing

Journal: bioRxiv

Article Title: Defining the chromatin-associated protein landscapes on Trypanosoma brucei repetitive elements using synthetic TALE proteins

doi: 10.1101/2025.04.22.649942

Figure Lengend Snippet: A. 177 repeat consensus sequence. Sequence that 177R-TALE-YFP was designed to bind is indicated. B. Distribution of 177R-TALE-YFP, TelR-TALE-YFP, YFP-TRF and 70R-TALE-YFP, at two intermediate/mini- chromosome telomeres determined by anti-GFP ChIP-seq. Anti-GFP ChIP-seq of 427 cells expressing no tagged protein is included as control. Diagrams below ChIP-seq profiles indicate the positions of 177bp repeats (red chevrons), 70bp repeats (green bars), VSG encoding genes (blue) and telomere (TTAGGG)n repeats (black chevrons) within Tb427VSG-671_unitig_Tb427v12:17,836-31,606 (31kb) and Tb427VSG-647_untig_Tb427v12 (10kb).

Article Snippet: Rabbit anti-GFP primary antibody (Thermo Fisher Scientific A-11122) was used at 1:500 dilution, and secondary Alexa fluor 568 goat antirabbit antibody (Thermo Fisher Scientific A-11036) was used at 1:1000 dilution.

Techniques: Sequencing, ChIP-sequencing, Expressing, Control

Journal: bioRxiv

Article Title: Defining the chromatin-associated protein landscapes on Trypanosoma brucei repetitive elements using synthetic TALE proteins

doi: 10.1101/2025.04.22.649942

Figure Lengend Snippet: A. Distribution of 177R-TALE, YFP-KKT2 and 147R-TALE over two intermediate/mini- chromosome telomeres determined by anti-GFP ChIP-seq. Anti-GFP ChIP-seq of T. brucei 427 cells expressing no tagged protein is included as control. Diagram below ChIP-seq profiles indicates the positions of 177bp repeats (red chevrons), 70bp repeats (green bars), VSG encoding genes (blue) and telomere (TTAGGG) n repeats (black chevrons) within Tb427VSG-671_unitig_Tb427v12:17,836-31,606 (31kb) and Tb427VSG-647_untig_Tb427v12 (10kb). B. Comparison of distribution of 177R-TALE, 147R-TALE and YFP-KKT2 over the chromosome 4 CIR147 centromere repeat array and adjacent unique sequences. Chr4:880,000-895,000 (15Kb) and Tb427VSG-671_unitig_Tb427v12:12,000-27,000 (31kb). Diagram below ChIP-seq indicates position of CIR147 repeats.

Article Snippet: Rabbit anti-GFP primary antibody (Thermo Fisher Scientific A-11122) was used at 1:500 dilution, and secondary Alexa fluor 568 goat antirabbit antibody (Thermo Fisher Scientific A-11036) was used at 1:1000 dilution.

Techniques: ChIP-sequencing, Expressing, Control, Comparison