rabbit anti adar1 primary antibody  (Cell Signaling Technology Inc)

Journal: bioRxiv

Article Title: Z-Form Stabilization By The Zα Domain Of Adar1p150 Has Subtle Effects On A-To-I Editing

doi: 10.1101/2025.06.02.657529

Figure Lengend Snippet: ( A ) The domain structure of the short (ADAR1p110) and long (ADAR1p150) isoforms of ADAR1 are shown. ADAR1p110 contains a deaminase domain which is responsible for ADAR1’s catalytic deamination activity, three double-stranded RNA binding domains (dsRBDs) which interact with the A-form structure of dsRNA, a Nuclear Localization Sequence (NLS) and a Zβ domain of unknown function. ADAR1p150 contains the same domain structure but has a ∼300 a.a. N-terminal extension which contains a Nuclear Export Sequence (NES) as well as a Zα domain. ( B ) ADAR1 deaminases adenosine to inosine in dsRNA, which replaces the amino group on the adenosine with a keto group and disrupts A-form helical structure at AU base pairs. ( C ) Cartoon model depicting editing of a dsRNA by ADAR1 and the different domains. ( D ) The Zα domain is able to stabilize the left-handed Z-conformation of dsDNA and dsRNA through key residues which stabilize the unique Z-form geometry.

Article Snippet: After blocking, the membrane was incubated overnight at 4°C with a rabbit anti-ADAR1 primary antibody (Cell Signaling, #14175) diluted 1:1000 in blocking buffer.

Techniques: Activity Assay, RNA Binding Assay, Sequencing

Journal: bioRxiv

Article Title: Z-Form Stabilization By The Zα Domain Of Adar1p150 Has Subtle Effects On A-To-I Editing

doi: 10.1101/2025.06.02.657529

Figure Lengend Snippet: ( A ) Domain architectures and point mutant locations for the different re-integrated ADAR1p150 mutant constructs. Red stars indicated mutation sites. ( B ) Western blot (left) and quantification (right) showing doxycycline-inducible expression levels of the re-integrated ADAR1p150 mutants relative to re-integrated wild-type ADAR1p150. Quantification was from three replicates, all of which are shown in Supplemental Figures 5 and 6. ( C ) mRNA expression levels in Transcripts Per Million (TPM, on the y-axis ) of the re-integrated ADAR1p150 constructs for each cell line from RNA-seq data. For each transgene, the detected expression value is indicated for each sample. ( D ) Principal Component Analysis (PCA) of gene expression of the wild-type HEK293T cells, ADAR1 KO and ADAR1p150 KO HEK293T cells, and the ADAR1p150 re-integrated cell lines.

Article Snippet: After blocking, the membrane was incubated overnight at 4°C with a rabbit anti-ADAR1 primary antibody (Cell Signaling, #14175) diluted 1:1000 in blocking buffer.

Techniques: Mutagenesis, Construct, Western Blot, Expressing, RNA Sequencing, Gene Expression

Journal: bioRxiv

Article Title: Z-Form Stabilization By The Zα Domain Of Adar1p150 Has Subtle Effects On A-To-I Editing

doi: 10.1101/2025.06.02.657529

Figure Lengend Snippet: ( A ) Immunofluorescence images of the wild-type, ADAR1 KO, ADAR1p150 KO, re-integrated ADAR1p150 (ADAR1p150*), the N173S mutant (ADAR1p150 N173S *), and the N173A,Y177A (ADAR1p150 N173A,Y177A *) double mutant cell lines. The red signal is of an ADAR1p150-specific rabbit monoclonal antibody visualized using an Alexa Fluor 594 nm secondary antibody. The green signal is from G3BP1 which was endogenously tagged with EGFP, and the blue signal is DAPI. ( B ) The percentage of the red signal intensity (corresponding to the anti-ADAR1p150 antibody) in the cytoplasm versus the nucleus of the cell.

Article Snippet: After blocking, the membrane was incubated overnight at 4°C with a rabbit anti-ADAR1 primary antibody (Cell Signaling, #14175) diluted 1:1000 in blocking buffer.

Techniques: Immunofluorescence, Mutagenesis

Journal: bioRxiv

Article Title: Z-Form Stabilization By The Zα Domain Of Adar1p150 Has Subtle Effects On A-To-I Editing

doi: 10.1101/2025.06.02.657529

Figure Lengend Snippet: ( A ) The structure of the Zα domain of ADAR1 bound to Z-RNA is shown (PDB: 2GXB) highlighting the water-mediated hydrogen bond between N173 and W195. ( B ) Domain structures of the ADAR1p150 N173S and ADAR1p150 N173S NES moved constructs. ( C ) Immunofluorescence images of the re-integrated wild-type ADAR1p150*, ADAR1p150 N173S , and ADAR1p150 N173S * NES moved cell lines. ( D ) percentage of Alexa Fluor 594 signal intensity (corresponding to anti-ADAR1p150 antibody staining) measured in the cytoplasm versus the nucleus of the cell for the ADAR1p150 N173S and ADAR1p150 N173S * NES moved cell lines.

Article Snippet: After blocking, the membrane was incubated overnight at 4°C with a rabbit anti-ADAR1 primary antibody (Cell Signaling, #14175) diluted 1:1000 in blocking buffer.

Techniques: Construct, Immunofluorescence, Staining

Journal: bioRxiv

Article Title: Z-Form Stabilization By The Zα Domain Of Adar1p150 Has Subtle Effects On A-To-I Editing

doi: 10.1101/2025.06.02.657529

Figure Lengend Snippet: ( A ) Domain structures of ADAR1p150 and ZBP1 are shown. ( B ) A cartoon depiction of the two potential models showing the effect of the Zα domain on ADAR1 function. In model 1, the Zα domain augments A-to-I editing broadly in a sequence-independent manner. In model 2, the Zα domain of ADAR1 competes with ZBP1 for binding to Z-form substrates in an editing independent manner, thereby inhibiting cell death pathways.

Article Snippet: After blocking, the membrane was incubated overnight at 4°C with a rabbit anti-ADAR1 primary antibody (Cell Signaling, #14175) diluted 1:1000 in blocking buffer.

Techniques: Sequencing, Binding Assay

Journal: Cancer medicine

Article Title: ADAR1 is a prognostic biomarker and is correlated with immune infiltration in lung adenocarcinoma.

doi: 10.1002/cam4.6044

Figure Lengend Snippet: FIGURE 1 Excessive expression of adenosine deaminase RNA-specific 1 (ADAR1) contributes to poor prognosis in non-small cell lung cancer (NSCLC). (A) Representative images of IHC staining with an ADAR1 antibody on lung adenocarcinoma (LUAD) and lung squamous carcinoma (LUSC) tumor tissues and their adjacent normal lung tissues. Scale bars: 200 μm. (B) Proportion of ADAR1high and ADAR1low cases of LUAD or LUSC. (C) ADAR1 mRNA expression levels in NSCLC, LUAD, and LUSC (***p < 0.001). (D) Overall survival curves based on ADAR1 mRNA expression in patients with NSCLC, LUSC, or LUAD.

Article Snippet: After that, the sections were incubated with 100 μL of anti- ADAR1 primary antibody (1:100, #81284, Cell Signaling Technology) overnight at 4°C.

Techniques: Expressing, Immunohistochemistry

Journal: Cancer medicine

Article Title: ADAR1 is a prognostic biomarker and is correlated with immune infiltration in lung adenocarcinoma.

doi: 10.1002/cam4.6044

Figure Lengend Snippet: FIGURE 2 Adenosine deaminase RNA-specific 1 (ADAR1) expression is associated with the clinicopathological characteristics and prognosis of patients with lung adenocarcinoma (LUAD). (A) Analysis of ADAR1 expression levels based on the clinical stages of The Cancer Genome Atlas-LUAD. (B) The expression levels of ADAR1 in stage I, stage II, stage III, stage IV, and normal tissues of patients with LUAD from the present cohort (n = 100). (C) Immunoreactivity score of ADAR1 expression in patients with LUAD at different stages. (D) K-M survival analysis of overall survival for LUAD patients according to the level of ADAR1 protein expression. (E) Univariate and (F) multivariate Cox analyses of ADAR1 in LUAD patients. (***p < 0.001, **p < 0.01, ns p > 0.5 nonsignificant).

Article Snippet: After that, the sections were incubated with 100 μL of anti- ADAR1 primary antibody (1:100, #81284, Cell Signaling Technology) overnight at 4°C.

Techniques: Expressing

Journal: Cancer medicine

Article Title: ADAR1 is a prognostic biomarker and is correlated with immune infiltration in lung adenocarcinoma.

doi: 10.1002/cam4.6044

Figure Lengend Snippet: FIGURE 3 Elevated adenosine deaminase RNA-specific 1 (ADAR1) expression is associated with lymph node metastasis (LNM) in lung adenocarcinoma (LUAD). ADAR1 expression level in LUAD with or without LNM (A) and their immunoreactive scores (B). Scale bars: 100 μm. ADAR1 expression level in primary LUAD or paired LNM lesions (C) and their immunoreactive scores (D). (mean ± SD, Student's t-test, **p < 0.01, *p < 0.05).

Article Snippet: After that, the sections were incubated with 100 μL of anti- ADAR1 primary antibody (1:100, #81284, Cell Signaling Technology) overnight at 4°C.

Techniques: Expressing

Journal: Cancer medicine

Article Title: ADAR1 is a prognostic biomarker and is correlated with immune infiltration in lung adenocarcinoma.

doi: 10.1002/cam4.6044

Figure Lengend Snippet: FIGURE 4 Correlation of adenosine deaminase RNA-specific 1 (ADAR1) with tumor immune infiltration in lung adenocarcinoma (LUAD). Correlation between ADAR1 and 22 tumor-infiltrating immune cells in LUAD samples as analyzed via cell-type identification by estimating relative subsets of known RNA transcripts. An absolute value of |Rho| > 0.1 and p < 0.05 implied that the ADAR1 expression level was significantly associated with the number of immune cells.

Article Snippet: After that, the sections were incubated with 100 μL of anti- ADAR1 primary antibody (1:100, #81284, Cell Signaling Technology) overnight at 4°C.

Techniques: Expressing

Journal: Cancer medicine

Article Title: ADAR1 is a prognostic biomarker and is correlated with immune infiltration in lung adenocarcinoma.

doi: 10.1002/cam4.6044

Figure Lengend Snippet: FIGURE 5 Adenosine deaminase RNA-specific 1 (ADAR1) is involved in immune cell infiltration in lung adenocarcinoma (LUAD) tissue. (A) Multiplex immunohistochemistry staining of LUAD sections of the high- (upper panel) or low- (lower panel) ADAR1 group. (B) Histograms displaying the quantitative data of CD4+, CD68+, CD20+, CD84+, CD86+, and CD206+ cells in the ADAR1-high and ADAR1- low groups. ADAR1+ (magenta), CD4+ T cells (CD4+, green), pan-macrophages (CD68+, yellow), CD8+ T cells (CD8+, red), B cells (CD20+, celeste), myeloid-derived suppressor cells (CD84+, white), M1 macrophages (CD86+, purple), and M2 macrophages (CD206+, orange). (mean ± SD, Student's t-test, ****p < 0.0001, ***p < 0.001, **p < 0.01, *p < 0.05, ns p > 0.05, not significant).

Article Snippet: After that, the sections were incubated with 100 μL of anti- ADAR1 primary antibody (1:100, #81284, Cell Signaling Technology) overnight at 4°C.

Techniques: Multiplex Assay, Immunohistochemistry, Staining, Derivative Assay