antibodies against ager  (R&D Systems)

Journal: Heliyon

Article Title: Human intestine and placenta exhibit tissue-specific expression of RAGE isoforms.

doi: 10.1016/j.heliyon.2023.e18247

Figure Lengend Snippet: Fig. 3. Schematic representation indicating immunogenic sequences within human full-length RAGE protein of the anti-RAGE antibodies tested in this study. Indicated domains: signal peptide (SP), variable immunoglobulin domain (IgV), two constant Ig domains (IgC1 and IgC2), trans membrane domain (TM) and cytoplasmic tail (CT).

Article Snippet: Antibodies against human RAGE (Fig. 3) were purchased from R&D Systems (AF1145; goat polyclonal IgG antibody raised against mouse myeloma cell line NS0-derived recombinant human RAGE Gln24-Ala344 and MAB11451; clone #176902; mouse monoclonal IgG2b antibody raised against mouse myeloma cell line NS0-derived recombinant human RAGE Gln24-Ala344), Santa Cruz (sc-80652; clone A11; mouse monoclonal IgG2a antibody raised against a truncated extracellular domain of RAGE of human origin) and Abnova (H00000177-D01P; rabbit polyclonal IgG antibody raised against a full-length human RAGE protein).

Techniques: Membrane

Journal: Heliyon

Article Title: Human intestine and placenta exhibit tissue-specific expression of RAGE isoforms.

doi: 10.1016/j.heliyon.2023.e18247

Figure Lengend Snippet: Fig. 5. RAGE immunoblotting analysis of human intestine. SDS-PAGE of lung lysate (1.5 μg/lane) as well as lysates from small intestine (SI) and colon (50 μg/lane) was performed under reducing conditions. Membranes were stained with three different anti-RAGE antibodies; SC A11 (A), AF1145 (B), and AB D01P (C). Membranes were also stained with the respective secondary antibodies only (negative control). GAPDH was used as a loading control. The uncropped images are shown in the supplementary material of the manuscript.

Article Snippet: Antibodies against human RAGE (Fig. 3) were purchased from R&D Systems (AF1145; goat polyclonal IgG antibody raised against mouse myeloma cell line NS0-derived recombinant human RAGE Gln24-Ala344 and MAB11451; clone #176902; mouse monoclonal IgG2b antibody raised against mouse myeloma cell line NS0-derived recombinant human RAGE Gln24-Ala344), Santa Cruz (sc-80652; clone A11; mouse monoclonal IgG2a antibody raised against a truncated extracellular domain of RAGE of human origin) and Abnova (H00000177-D01P; rabbit polyclonal IgG antibody raised against a full-length human RAGE protein).

Techniques: Western Blot, SDS Page, Staining, Negative Control, Control

Journal: Heliyon

Article Title: Human intestine and placenta exhibit tissue-specific expression of RAGE isoforms.

doi: 10.1016/j.heliyon.2023.e18247

Figure Lengend Snippet: Fig. 4. (A) RAGE immunoblotting analysis of human lung lysate. SDS-PAGE of lung lysate (1 μg/lane; PNGase F -) and deglycosylated lung lysate (1 μg/lane; PNGase F +) was performed under reducing conditions. Membranes were stained with four different anti-RAGE antibodies. Membranes were also stained with the respective secondary antibodies only (negative control). GAPDH was used as a loading control. (B) RAGE immunoblotting analysis of RAGE overexpressing (+) and control (−) HEK293T cells. SDS-PAGE was performed under reducing conditions. For WB analysis membranes were stained with anti-FLAG and anti-RAGE (AB D01P) antibodies. The uncropped images are shown in the supplementary material of the manuscript.

Article Snippet: Antibodies against human RAGE (Fig. 3) were purchased from R&D Systems (AF1145; goat polyclonal IgG antibody raised against mouse myeloma cell line NS0-derived recombinant human RAGE Gln24-Ala344 and MAB11451; clone #176902; mouse monoclonal IgG2b antibody raised against mouse myeloma cell line NS0-derived recombinant human RAGE Gln24-Ala344), Santa Cruz (sc-80652; clone A11; mouse monoclonal IgG2a antibody raised against a truncated extracellular domain of RAGE of human origin) and Abnova (H00000177-D01P; rabbit polyclonal IgG antibody raised against a full-length human RAGE protein).

Techniques: Western Blot, SDS Page, Staining, Negative Control, Control

Journal: Heliyon

Article Title: Human intestine and placenta exhibit tissue-specific expression of RAGE isoforms.

doi: 10.1016/j.heliyon.2023.e18247

Figure Lengend Snippet: Fig. 6. RAGE immunoblotting analysis of human placenta derived from different healthy (H) as well as GDM-, FGR- and PE-affected pregnancies. SDS-PAGE of lung lysate (1 μg/lane) as well as lysates from the placentas (25 μg/lane) was performed under reducing conditions. For WB analysis membranes were stained with three different anti-RAGE antibodies; SC A11 (A), AF1145 (B), and AB D01P (C). Membranes were also stained with the respective secondary antibodies only (negative control). Ponceau S staining was used as a loading control. The uncropped images are shown in the supplementary material of the manuscript.

Article Snippet: Antibodies against human RAGE (Fig. 3) were purchased from R&D Systems (AF1145; goat polyclonal IgG antibody raised against mouse myeloma cell line NS0-derived recombinant human RAGE Gln24-Ala344 and MAB11451; clone #176902; mouse monoclonal IgG2b antibody raised against mouse myeloma cell line NS0-derived recombinant human RAGE Gln24-Ala344), Santa Cruz (sc-80652; clone A11; mouse monoclonal IgG2a antibody raised against a truncated extracellular domain of RAGE of human origin) and Abnova (H00000177-D01P; rabbit polyclonal IgG antibody raised against a full-length human RAGE protein).

Techniques: Western Blot, Derivative Assay, SDS Page, Staining, Negative Control, Control

Journal: Heliyon

Article Title: Human intestine and placenta exhibit tissue-specific expression of RAGE isoforms.

doi: 10.1016/j.heliyon.2023.e18247

Figure Lengend Snippet: Fig. 7. RAGE immunoblotting analysis of human epithelial cell lines. SDS-PAGE of JE6-1 RAGE, JE6-1, Caco-2, FHs 74 Int and BeWo cells (25 μg/ lane) was performed under reducing conditions. For WB analysis membranes were stained with three different anti-RAGE antibodies; SC A11 (A), AF1145 (B), and AB D01P (C). To identify unspecific binding, membranes were also stained with the respective secondary antibodies only (negative control). GAPDH staining was used as a loading control. The uncropped images are shown in the supplementary material of the manuscript.

Article Snippet: Antibodies against human RAGE (Fig. 3) were purchased from R&D Systems (AF1145; goat polyclonal IgG antibody raised against mouse myeloma cell line NS0-derived recombinant human RAGE Gln24-Ala344 and MAB11451; clone #176902; mouse monoclonal IgG2b antibody raised against mouse myeloma cell line NS0-derived recombinant human RAGE Gln24-Ala344), Santa Cruz (sc-80652; clone A11; mouse monoclonal IgG2a antibody raised against a truncated extracellular domain of RAGE of human origin) and Abnova (H00000177-D01P; rabbit polyclonal IgG antibody raised against a full-length human RAGE protein).

Techniques: Western Blot, SDS Page, Staining, Binding Assay, Negative Control, Control

Journal: Heliyon

Article Title: Human intestine and placenta exhibit tissue-specific expression of RAGE isoforms.

doi: 10.1016/j.heliyon.2023.e18247

Figure Lengend Snippet: Fig. 8. RAGE subcellular localization in human cell lines. (A) JE6-1 cells overexpressing full-length RAGE as well as wild-type control cells were stained with four different RAGE-specific antibodies (SC A11, AF1145, AB D01P and MAB11451). Nuclei were stained with DRAQ5. Scale bar: 25 μm. (B) RAGE staining of JE6-1 cells and intestinal epithelial cells Caco-2 using antibody SC A11 and prolonged exposure times. Controls were stained with the respective secondary antibody only. Nuclei were stained with DRAQ5. Scale bar: 25 μm.

Article Snippet: Antibodies against human RAGE (Fig. 3) were purchased from R&D Systems (AF1145; goat polyclonal IgG antibody raised against mouse myeloma cell line NS0-derived recombinant human RAGE Gln24-Ala344 and MAB11451; clone #176902; mouse monoclonal IgG2b antibody raised against mouse myeloma cell line NS0-derived recombinant human RAGE Gln24-Ala344), Santa Cruz (sc-80652; clone A11; mouse monoclonal IgG2a antibody raised against a truncated extracellular domain of RAGE of human origin) and Abnova (H00000177-D01P; rabbit polyclonal IgG antibody raised against a full-length human RAGE protein).

Techniques: Control, Staining

Journal: International journal of biological sciences

Article Title: Soluble Receptor for Advanced Glycation End-products regulates age-associated Cardiac Fibrosis.

doi: 10.7150/ijbs.56379

Figure Lengend Snippet: Figure 2. RAGE deficiency promotes the expression of HF marker genes and accelerates age-associated cardiac fibrosis. (A) Relative expression of BNP (Nppb) and ANKRD1 (Ankrd1) genes. Young WT n=4, Young Rage-/- n=4-5, Middle-age (MA) WT n=4, MA Rage-/- n=4, Old WT n=5, Old Rage-/- n=6. (B) (Left panel) Representative images of LV sections stained with an antibody against Collagen I. Bar, 20 µm. (Right panel) Quantification of Collagen I signal. Young WT n=5, Young Rage-/- n=5, MA WT n=4, MA Rage-/- n=4, Old WT n=4, Old Rage-/- n=5. (C) (Left panel) Representative images of LV sections stained with Masson’s trichrome. Bar, 20 µm. (Right panel) Quantification of Masson’s trichrome signal. Young WT n=5, Young Rage-/- n=5, MA WT n=4, MA Rage-/- n=4, Old WT n=4, Old Rage-/- n=5. (D) (Left panel) Representative images of LV sections stained with an antibody anti-α-SMA. Bar, 20 µm. (Right panel) Quantification of α-SMA signal. Young WT n=5, Young Rage-/- n=5, MA WT n=4, MA Rage-/- n=4, Old WT n=4, Old Rage-/- n=5. Each dot represents a mouse; mean ± SD are shown (*, P<0.05; **, P<0.01; ***, P<0.001; 2-way ANOVA plus Bonferroni post-hoc test for multiple comparisons).

Article Snippet: Membranes were probed with antibodies against RAGE (RAGE N-term, 1 μg/ml; #AF1145, R&D Systems Inc.), Collagen I (1 μg/ml; #ab34710, Abcam), Collagen III (0.5 μg/ml; #ab7778, Abcam), α-SMA (0.2 μg/ml; #ab5694; Abcam), TGFbetaR1 (1 μg/ml, ABF17-I, EMD Millipore, USA), βTubulin (1 μg/ml; #T-6199, Sigma-Aldrich), βactin (1:1000; #A5441, Sigma) or GAPDH (0.2 μg/ml, sc-25778, Santa Cruz Biotechnology, Dallas, Texas, USA).

Techniques: Expressing, Marker, Staining

Journal: International journal of biological sciences

Article Title: Soluble Receptor for Advanced Glycation End-products regulates age-associated Cardiac Fibrosis.

doi: 10.7150/ijbs.56379

Figure Lengend Snippet: Figure 3. RAGE deficiency induces age-associated TGF-β signaling. (A) Relative expression of Tgfb1gene. Young WT n=4, Young Rage-/- n=4-5, Middle-age (MA) WT n=4, MA Rage-/- n=4, Old WT n=5, Old Rage-/- n=6. (B) (Left panel) Representative images of LV sections stained with an anti-Smad2-3 antibody. Bar, 20 µm. (Right panel) Quantification of Smad2/3 signal. Young WT n=5, Young Rage-/- n=5, MA WT n=4, MA Rage-/- n=4, Old WT n=4, Old Rage-/- n=5. (C) (Left panel) Representative images of LV sections stained with an anti-TFGbetaR1 antibody. Bar, 20 µm. (Right panel) Quantification of TFGbetaR1 signal. Young WT n=5, Young Rage-/- n=5, MA WT n=4, MA Rage-/- n=4, Old WT n=4, Old Rage-/- n=5. Each dot represents a mouse; mean ± SD are shown (*, P<0.05; **, P<0.01; ***, P<0.001; 2-way ANOVA plus Bonferroni post-hoc test for multiple comparisons).

Article Snippet: Membranes were probed with antibodies against RAGE (RAGE N-term, 1 μg/ml; #AF1145, R&D Systems Inc.), Collagen I (1 μg/ml; #ab34710, Abcam), Collagen III (0.5 μg/ml; #ab7778, Abcam), α-SMA (0.2 μg/ml; #ab5694; Abcam), TGFbetaR1 (1 μg/ml, ABF17-I, EMD Millipore, USA), βTubulin (1 μg/ml; #T-6199, Sigma-Aldrich), βactin (1:1000; #A5441, Sigma) or GAPDH (0.2 μg/ml, sc-25778, Santa Cruz Biotechnology, Dallas, Texas, USA).

Techniques: Expressing, Staining

Journal: International journal of biological sciences

Article Title: Soluble Receptor for Advanced Glycation End-products regulates age-associated Cardiac Fibrosis.

doi: 10.7150/ijbs.56379

Figure Lengend Snippet: Figure 4. Circulating sRAGE decreases with age in mice. (A, B) RAGE isoforms are not detectable in LV tissue. (A) mRNA levels of mRAGE or mRAGE+mRAGE_v4 isoforms in LV or lung (positive control) of indicated WT age groups. (B) Forty µg of LV protein lysate of the indicated age groups of WT mice were probed with an antibody α-RAGE (RAGE N-term). The same amount of LV from a Young Rage-/- mouse (KO) was used as negative control, and 10 µg of lung protein lysate of a WT mouse (L) as positive control for RAGE expression. β-tubulin was used as loading control (n=3-4 per group). (C) Quantification of sRAGE in the serum of the indicated age groups of WT mice; Young n=17, MA n=17 and Old n=18. (D) (Left panel) Ten µg of lung protein lysate of the indicated age groups of WT mice were probed with an antibody against RAGE (RAGE N-term) and a long and a short exposure are shown. Ten µg of lung protein lysate from a Young Rage-/- mouse (KO) was used as negative control. α-actin was used as loading control. (Right panel) Quantification of RAGE isoforms in the lung protein lysate of the indicated age groups of WT mice (n=3-5 per group). Each dot represents a mouse; mean ± SD are shown (*, P<0.05; **, P<0.01; ***, P<0.001; 1-way ANOVA plus Bonferroni post-hoc test for multiple comparisons).

Article Snippet: Membranes were probed with antibodies against RAGE (RAGE N-term, 1 μg/ml; #AF1145, R&D Systems Inc.), Collagen I (1 μg/ml; #ab34710, Abcam), Collagen III (0.5 μg/ml; #ab7778, Abcam), α-SMA (0.2 μg/ml; #ab5694; Abcam), TGFbetaR1 (1 μg/ml, ABF17-I, EMD Millipore, USA), βTubulin (1 μg/ml; #T-6199, Sigma-Aldrich), βactin (1:1000; #A5441, Sigma) or GAPDH (0.2 μg/ml, sc-25778, Santa Cruz Biotechnology, Dallas, Texas, USA).

Techniques: Positive Control, Negative Control, Expressing, Control