antibodies avivasysbio  (Alomone Labs)

Journal: Redox Report : Communications in Free Radical Research

Article Title: ASIC1a Promotes nucleus pulposus derived stem cells apoptosis through modulation of SIRT3-dependent mitochondrial redox homeostasis in intervertebral disc degeneration

doi: 10.1080/13510002.2025.2504120

Figure Lengend Snippet: Changes of degenerated intervertebral disc tissue in human and rat. (A)Representative MRI T2-weighted images of human IVD of grade II and grade IV degeneration. (B) General view (Scale bar: 1 cm), H&E (Scale bar: 25 μM) and Alcian blue stainings (Scale bar: 25 μM) of human nucleus pulposus of grade II and grade IV degeneration. (C) IHC staining of ASIC1a and c-caspase3 in human nucleus pulposus sections. Scale bar: 40 μM. (D) IHC staining of ASIC1a and c-caspase3 in rats nucleus pulposus sections. Scale bar: 20 μM. H&E, hematoxylin and eosin; IVD, intervertebral disc.

Article Snippet: The primary antibodies utilized included ASIC1a (Huaan ER1803-33, 1:1000), SIRT1 (Huaan ER130811 , 1:1000), SIRT2 (Huaan ET1611-72, 1:1000), SIRT3 (Immunoway YT4304, 1:1000), SIRT4 (Proteintech 66543-ig, 1:20000), SIRT5 (Huaan ET1701-6, 1:1000), SIRT6 (Huaan ET1701-29, 1:1000), SIRT7 (Huaan EM1707-09, 1:1000), BAX (Proteintech 50599-2-ig, 1:5000), Bcl-2 (Proteintech 26593-1-AP, 1:2000), Active Caspase-3 (Huaan ET1602-47, 1:1000), Cytochrome C (Huaan ET1610-16, 1:1000), SOD2 (Immunoway YT5575, 1:1000), AMPK (Huaan ET1608-40, 1:4000), p-AMPK (Huaan ET1612-72, 1:1000), PGC-1α (Huaan ET1702-96, 1:1000), VDAC (Huaan ET1601-20, 1:10000), and Beta Actin (Proteintech 20536-1-AP, 1:10000).

Techniques: Immunohistochemistry

Journal: Redox Report : Communications in Free Radical Research

Article Title: ASIC1a Promotes nucleus pulposus derived stem cells apoptosis through modulation of SIRT3-dependent mitochondrial redox homeostasis in intervertebral disc degeneration

doi: 10.1080/13510002.2025.2504120

Figure Lengend Snippet: Acid treatment inhibited human NPSCs proliferation, promoted mitochondrial pathway apoptosis and ASIC1a expression. The human NPSCs were treated with acid medium of pH 7.4, 7.1, 6.8, 6.5 for 0–36 h. (A) Cell viability was examined by the absorbance of CCK-8, the results are expressed as OD at 450 nm. (B-E) Representative western blotting assay and quantitation of the level of ASIC1a, the ratio of Bcl-2/Bax and cleaved caspase 3. (F, G) TUNEL staining to detect human NPSCs apoptosis induced by acidosis treatment and analyzed using fluorescence microscope. Cell nuclei are stained by DAPI. Scale bar: 50 μM. (H, I) JC-1 staining to detect the level of MMP in acid-treated human NPSCs and analyzed using fluorescence microscope. Scale bar: 50 μM. (J, K) Representative western blotting assay and quantitation of the level of the ratio of cytochrome c (Cyt-c) in mitochondrial and cytoplasmic extracts. (L) Cell apoptosis was measured by Annexin V/PI staining under flow cytometry analysis. Data are mean ± SD, ns: no statistical significance, ** indicating P ≤ 0.01, *** indicating P ≤ 0.001. NPSC, nucleus pulposus-derived stem cell.

Article Snippet: The primary antibodies utilized included ASIC1a (Huaan ER1803-33, 1:1000), SIRT1 (Huaan ER130811 , 1:1000), SIRT2 (Huaan ET1611-72, 1:1000), SIRT3 (Immunoway YT4304, 1:1000), SIRT4 (Proteintech 66543-ig, 1:20000), SIRT5 (Huaan ET1701-6, 1:1000), SIRT6 (Huaan ET1701-29, 1:1000), SIRT7 (Huaan EM1707-09, 1:1000), BAX (Proteintech 50599-2-ig, 1:5000), Bcl-2 (Proteintech 26593-1-AP, 1:2000), Active Caspase-3 (Huaan ET1602-47, 1:1000), Cytochrome C (Huaan ET1610-16, 1:1000), SOD2 (Immunoway YT5575, 1:1000), AMPK (Huaan ET1608-40, 1:4000), p-AMPK (Huaan ET1612-72, 1:1000), PGC-1α (Huaan ET1702-96, 1:1000), VDAC (Huaan ET1601-20, 1:10000), and Beta Actin (Proteintech 20536-1-AP, 1:10000).

Techniques: Expressing, CCK-8 Assay, Western Blot, Quantitation Assay, TUNEL Assay, Staining, Fluorescence, Microscopy, Flow Cytometry, Derivative Assay

Journal: Redox Report : Communications in Free Radical Research

Article Title: ASIC1a Promotes nucleus pulposus derived stem cells apoptosis through modulation of SIRT3-dependent mitochondrial redox homeostasis in intervertebral disc degeneration

doi: 10.1080/13510002.2025.2504120

Figure Lengend Snippet: Knockdown and overexpression of ASIC1a affected acidosis-treated human NPSC apoptosis via mitochondrial pathway. (A, B) Representative western blotting assay and quantitation of the level of ASIC1a. (C-F) Representative western blotting assay and quantitation of the level of ASIC1a, cleaved caspase 3, and the ratio of Bcl-2/Bax. (G, H) TUNEL staining to detect human NPSCs apoptosis induced by acidosis treatment and analyzed using fluorescence microscope. Cell nuclei are stained by DAPI. Scale bar: 50 μM. (I, J) JC-1 staining to detect the level of MMP in acidosis-treated human NPSCs and analyzed using fluorescence microscope. Scale bar: 50 μM. (K, L) Representative western blotting assay and quantitation of the level of the ratio of cytochrome c (Cyt-c) in mitochondrial and cytoplasmic extracts. (M, N) Representative western blotting assay and quantitation of the level of ASIC1a. (O-R) Representative western blotting assay and quantitation of the level of ASIC1a, cleaved caspase 3, and the ratio of Bcl-2/Bax. (S, T) TUNEL staining to detect human NPSCs apoptosis induced by acidosis treatment and analyzed using fluorescence microscope. Cell nuclei are stained by DAPI. Scale bar: 50 μM. (U, V) JC-1 staining to detect the level of MMP in acidosis-treated human NPSCs and analyzed using fluorescence microscope. Scale bar: 50 μM. (W, X) Representative western blotting assay and quantitation of the level of the ratio of cytochrome c (Cyt-c) in mitochondrial and cytoplasmic extracts. (Y) Cell apoptosis was measured by Annexin V/PI staining under flow cytometry analysis. Data are mean ± SD, * indicating P ≤ 0.05, ** indicating P ≤ 0.01, *** indicating P ≤ 0.001. NPSC, nucleus pulposus-derived stem cell.

Article Snippet: The primary antibodies utilized included ASIC1a (Huaan ER1803-33, 1:1000), SIRT1 (Huaan ER130811 , 1:1000), SIRT2 (Huaan ET1611-72, 1:1000), SIRT3 (Immunoway YT4304, 1:1000), SIRT4 (Proteintech 66543-ig, 1:20000), SIRT5 (Huaan ET1701-6, 1:1000), SIRT6 (Huaan ET1701-29, 1:1000), SIRT7 (Huaan EM1707-09, 1:1000), BAX (Proteintech 50599-2-ig, 1:5000), Bcl-2 (Proteintech 26593-1-AP, 1:2000), Active Caspase-3 (Huaan ET1602-47, 1:1000), Cytochrome C (Huaan ET1610-16, 1:1000), SOD2 (Immunoway YT5575, 1:1000), AMPK (Huaan ET1608-40, 1:4000), p-AMPK (Huaan ET1612-72, 1:1000), PGC-1α (Huaan ET1702-96, 1:1000), VDAC (Huaan ET1601-20, 1:10000), and Beta Actin (Proteintech 20536-1-AP, 1:10000).

Techniques: Knockdown, Over Expression, Western Blot, Quantitation Assay, TUNEL Assay, Staining, Fluorescence, Microscopy, Flow Cytometry, Derivative Assay

Journal: Redox Report : Communications in Free Radical Research

Article Title: ASIC1a Promotes nucleus pulposus derived stem cells apoptosis through modulation of SIRT3-dependent mitochondrial redox homeostasis in intervertebral disc degeneration

doi: 10.1080/13510002.2025.2504120

Figure Lengend Snippet: MitoTEMPO pre-treatment alleviated ASIC1a overexpression-induced mitochondrial oxidative stress and apoptosis in acidosis-treated human NPSCs. (A, B) Representative fluorescent images with DCFH-DA (green) staining. Scale bar: 100 μM. (C, D) Representative fluorescence images with MitoSOX (red) and MitoTracker (green) double – staining. Cell nuclei are stained by DAPI. Scale bar: 50 μM. (E, F) TUNEL staining to detect human NPSCs apoptosis induced by acidosis treatment and analyzed using fluorescence microscope. Cell nuclei are stained by DAPI. Scale bar: 50 μM. (G, H) JC-1 staining to detect the level of MMP in acidosis-treated human NPSCs and analyzed using fluorescence microscope. Scale bar: 50 μM. (I-K) Representative western blotting assay and quantitation of the level of cleaved caspase 3 and the ratio of Bcl-2/Bax. (L, M) Representative western blotting assay and quantitation of the level of the ratio of cytochrome c (Cyt-c) in mitochondrial and cytoplasmic extracts. Data are mean ± SD, * indicating P ≤ 0.05, ** indicating P ≤ 0.01, *** indicating P ≤ 0.001. NPSC, nucleus pulposus-derived stem cell.

Article Snippet: The primary antibodies utilized included ASIC1a (Huaan ER1803-33, 1:1000), SIRT1 (Huaan ER130811 , 1:1000), SIRT2 (Huaan ET1611-72, 1:1000), SIRT3 (Immunoway YT4304, 1:1000), SIRT4 (Proteintech 66543-ig, 1:20000), SIRT5 (Huaan ET1701-6, 1:1000), SIRT6 (Huaan ET1701-29, 1:1000), SIRT7 (Huaan EM1707-09, 1:1000), BAX (Proteintech 50599-2-ig, 1:5000), Bcl-2 (Proteintech 26593-1-AP, 1:2000), Active Caspase-3 (Huaan ET1602-47, 1:1000), Cytochrome C (Huaan ET1610-16, 1:1000), SOD2 (Immunoway YT5575, 1:1000), AMPK (Huaan ET1608-40, 1:4000), p-AMPK (Huaan ET1612-72, 1:1000), PGC-1α (Huaan ET1702-96, 1:1000), VDAC (Huaan ET1601-20, 1:10000), and Beta Actin (Proteintech 20536-1-AP, 1:10000).

Techniques: Over Expression, Staining, Fluorescence, Double Staining, TUNEL Assay, Microscopy, Western Blot, Quantitation Assay, Derivative Assay

Journal: Redox Report : Communications in Free Radical Research

Article Title: ASIC1a Promotes nucleus pulposus derived stem cells apoptosis through modulation of SIRT3-dependent mitochondrial redox homeostasis in intervertebral disc degeneration

doi: 10.1080/13510002.2025.2504120

Figure Lengend Snippet: The content and correlation of ASIC1a and SIRT3 in NP with different grades of degeneration. Twenty NP tissue specimens with different degenerative grades were used to analyze AISC1a and SIRT3 level by western blotting analysis. (A, B) Representative western blotting assay and quantitation of the levels of SIRT1-7. (C,D) Representative western blotting assay and quantitation of the levels of ASIC1a and SIRT3. (E) Correlation analysis between ASIC1a content and Pfirrmann grade was performed. (F) Correlation analysis between SIRT3 content and Pfirrmann grade was performed. (G) Correlation analysis between SIRT3 content and ASIC1a content. Data are mean ± SD, ns: no statistical significance, * indicating P ≤ 0.05, ** indicating P ≤ 0.01, *** indicating P ≤ 0.001. NPSC, nucleus pulposus-derived stem cell.

Article Snippet: The primary antibodies utilized included ASIC1a (Huaan ER1803-33, 1:1000), SIRT1 (Huaan ER130811 , 1:1000), SIRT2 (Huaan ET1611-72, 1:1000), SIRT3 (Immunoway YT4304, 1:1000), SIRT4 (Proteintech 66543-ig, 1:20000), SIRT5 (Huaan ET1701-6, 1:1000), SIRT6 (Huaan ET1701-29, 1:1000), SIRT7 (Huaan EM1707-09, 1:1000), BAX (Proteintech 50599-2-ig, 1:5000), Bcl-2 (Proteintech 26593-1-AP, 1:2000), Active Caspase-3 (Huaan ET1602-47, 1:1000), Cytochrome C (Huaan ET1610-16, 1:1000), SOD2 (Immunoway YT5575, 1:1000), AMPK (Huaan ET1608-40, 1:4000), p-AMPK (Huaan ET1612-72, 1:1000), PGC-1α (Huaan ET1702-96, 1:1000), VDAC (Huaan ET1601-20, 1:10000), and Beta Actin (Proteintech 20536-1-AP, 1:10000).

Techniques: Western Blot, Quantitation Assay, Derivative Assay

Journal: Redox Report : Communications in Free Radical Research

Article Title: ASIC1a Promotes nucleus pulposus derived stem cells apoptosis through modulation of SIRT3-dependent mitochondrial redox homeostasis in intervertebral disc degeneration

doi: 10.1080/13510002.2025.2504120

Figure Lengend Snippet: ASIC1a disturbed the mitochondrial redox homeostasis and increased apoptosis by regulating SIRT3. Human NPSCs were transfected with negative control siRNA (si-Control) or siRNA (si-ASIC1a or si-SIRT3), negative control plasmid (vector) or ASIC1a plasmid (OE-ASIC1a), and then treated with or without acidosis. (A-D) Representative western blotting assay and quantitation of the levels of SIRT3 and SOD2. (E-I) Representative western blotting assay and quantitation of the levels of SIRT3, SOD2, cleaved caspase 3, and the ratio of Bcl-2/Bax. (J, K) Representative fluorescent images with DCFH-DA (green) staining. Scale bar: 100 μM. (L, M) Representative fluorescence images with MitoSOX (red) and MitoTracker (green) double – staining. Cell nuclei are stained by DAPI. Scale bar: 50 μM. (N, O) TUNEL staining to detect human NPSCs apoptosis induced by acidosis treatment and analyzed using fluorescence microscope. Cell nuclei are stained by DAPI. Scale bar: 50 μM. (P, Q) JC-1 staining to detect the level of MMP in acidosis-treated human NPSCs and analyzed using fluorescence microscope. Scale bar: 50 μM. (R, S) Representative western blotting assay and quantitation of the level of the ratio of cytochrome c (Cyt-c) in mitochondrial and cytoplasmic extracts. Data are mean ± SD, * indicating P ≤ 0.05, ** indicating P ≤ 0.01, *** indicating P ≤ 0.001. NPSC, nucleus pulposus-derived stem cell.

Article Snippet: The primary antibodies utilized included ASIC1a (Huaan ER1803-33, 1:1000), SIRT1 (Huaan ER130811 , 1:1000), SIRT2 (Huaan ET1611-72, 1:1000), SIRT3 (Immunoway YT4304, 1:1000), SIRT4 (Proteintech 66543-ig, 1:20000), SIRT5 (Huaan ET1701-6, 1:1000), SIRT6 (Huaan ET1701-29, 1:1000), SIRT7 (Huaan EM1707-09, 1:1000), BAX (Proteintech 50599-2-ig, 1:5000), Bcl-2 (Proteintech 26593-1-AP, 1:2000), Active Caspase-3 (Huaan ET1602-47, 1:1000), Cytochrome C (Huaan ET1610-16, 1:1000), SOD2 (Immunoway YT5575, 1:1000), AMPK (Huaan ET1608-40, 1:4000), p-AMPK (Huaan ET1612-72, 1:1000), PGC-1α (Huaan ET1702-96, 1:1000), VDAC (Huaan ET1601-20, 1:10000), and Beta Actin (Proteintech 20536-1-AP, 1:10000).

Techniques: Transfection, Negative Control, Control, Plasmid Preparation, Western Blot, Quantitation Assay, Staining, Fluorescence, Double Staining, TUNEL Assay, Microscopy, Derivative Assay

Journal: Redox Report : Communications in Free Radical Research

Article Title: ASIC1a Promotes nucleus pulposus derived stem cells apoptosis through modulation of SIRT3-dependent mitochondrial redox homeostasis in intervertebral disc degeneration

doi: 10.1080/13510002.2025.2504120

Figure Lengend Snippet: ASIC1a inhibited the expression of SIRT3 by regulating AMPK/PGC-1α pathway. (A, B) Representative western blotting assay and quantitation of the levels of p-AMPK, AMPK, PGC-1α and SIRT3 in siRNA transfected NPSCs treated with or without acidosis. (C, D) Representative western blotting assay and quantitation of the levels of p-AMPK, AMPK, PGC-1α and SIRT3 in siRNA transfected NPSCs treated with acidosis in the presence or absence of Compound C exposure at 2.5 μM for 4 h. (E, F) Representative western blotting assay and quantitation of the levels of PGC-1α and SIRT3 in siRNA transfected NPSCs treated with acidosis. Data are mean ± SD, * indicating P ≤ 0.05, ** indicating P ≤ 0.01, *** indicating P ≤ 0.001. NPSC, nucleus pulposus-derived stem cell.

Article Snippet: The primary antibodies utilized included ASIC1a (Huaan ER1803-33, 1:1000), SIRT1 (Huaan ER130811 , 1:1000), SIRT2 (Huaan ET1611-72, 1:1000), SIRT3 (Immunoway YT4304, 1:1000), SIRT4 (Proteintech 66543-ig, 1:20000), SIRT5 (Huaan ET1701-6, 1:1000), SIRT6 (Huaan ET1701-29, 1:1000), SIRT7 (Huaan EM1707-09, 1:1000), BAX (Proteintech 50599-2-ig, 1:5000), Bcl-2 (Proteintech 26593-1-AP, 1:2000), Active Caspase-3 (Huaan ET1602-47, 1:1000), Cytochrome C (Huaan ET1610-16, 1:1000), SOD2 (Immunoway YT5575, 1:1000), AMPK (Huaan ET1608-40, 1:4000), p-AMPK (Huaan ET1612-72, 1:1000), PGC-1α (Huaan ET1702-96, 1:1000), VDAC (Huaan ET1601-20, 1:10000), and Beta Actin (Proteintech 20536-1-AP, 1:10000).

Techniques: Expressing, Western Blot, Quantitation Assay, Transfection, Derivative Assay