Human Aldehyde Dehydrogenase 1‑A1/
ALDH1A1 Antibody

Aldehyde Dehydrogenase 1‑A1/ALDH1A1 in A549 Human Cell Line. Aldehyde Dehydrogenase 1-A1/ALDH1A1 was detected in immersion fixed A549 human lung carcinoma cell line using Mouse Anti-Human Aldehyde Dehydrogenase 1-A1/ALDH1A1 Monoclonal Antibody (Catalog # MAB5869) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Aldehyde Dehydrogenase 1‑A1/ALDH1A1 in Human Brain. Aldehyde Dehydrogenase 1-A1/ALDH1A1 was detected in immersion fixed paraffin-embedded sections of human brain (substantia nigra) using Mouse Anti-Human Aldehyde Dehydrogenase 1-A1/ALDH1A1 Monoclonal Antibody (Catalog # MAB5869) at 5 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS002) and counterstained with hematoxylin (blue). Specific staining was localized to neurons. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Detection of Human Aldehyde Dehydrogenase 1‑A1/ALDH1A1 by Simple Western^TM. Simple Western lane view shows lysates of human liver tissue, loaded at 0.2 mg/mL. A specific band was detected for Aldehyde Dehydrogenase 1‑A1/ALDH1A1 at approximately 58 kDa (as indicated) using 20 µg/mL of Mouse Anti-Human Aldehyde Dehydrogenase 1‑A1/ALDH1A1 Monoclonal Antibody (Catalog # MAB5869). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Detection of Human, Mouse, and Rat Aldehyde Dehydrogenase 1‑A1/ALDH1A1 by Western Blot. Western blot shows lysates of human liver tissue, mouse liver tissue, and rat lung tissue. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human Aldehyde Dehydrogenase 1-A1/ALDH1A1 Monoclonal Antibody (Catalog # MAB5869) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for Aldehyde Dehydrogenase 1-A1/ ALDH1A1 at approximately 54 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of Human Aldehyde Dehydrogenase 1-A1/ALDH1A1 by Western Blot HTRA2 overexpression inhibitsDCIS.com breast cancer cell growth and invasion. (A–F) DCIS.com cells expressing pHAGE control or HTRA2 were analyzed for (A) ALDH1A1 and HTRA2 expression by immunoblot, (B) spheroid size in 3D cultures, (C) PCNA expression, (D) transwell invasion, (E) E-cadherin or (F) TWIST1 expression. Mean number of spheroids analyzed per group±s.e.m.: pHAGE, 325±25; pHAGE±CCL2, 358±33; HTRA2-OE, 343±17; HTRA2-OE±CCL2, 316±44. Protein levels in immunoblots were measured by densitometry. Expression levels were normalized to control group. Representative blots of three experiments are shown. Spheroid size and invasion index were normalized to spheroid number. Experimental groups were plated in triplicate. Experiments were performed three times. Two slides containing three sections each were subject to immunostaining. Statistical analysis was performed using two-tailed t-test (A) or one-way ANOVA with Bonferroni post-hoc comparison (B–F). *P<0.05, **P<0.01, ***P<0.001; n.s, not significant. Mean±s.e.m. is shown. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31208996), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human Aldehyde Dehydrogenase 1-A1/ALDH1A1 by Western Blot HTRA2 overexpression orknockdown affects CCR2-H SUM225 and CCR2-KO DCIS.com breast cancer cell growth and invasion. (A–D) CCR2-H SUM225 cells expressing vehicle control (pHAGE) or HTRA2 (HTRA2-OE) were analyzed for HTRA2 expression by immunoblot (A), spheroid size in 3D cultures (B), PCNA expression (C) or invasion index (D). Mean number of spheroids analyzed per group±s.e.m.: pHAGE, 269±63; HTRA2-OE, 227±28. (E–H) CCR2-KO DCIS.com cells were transduced with lentivirus expressing Con or HTRA2 shRNAs (HTRA2-KD#2, HTRA2-KD#3) and analyzed for HTRA2 expression by immunoblot (E), spheroid size in Matrigel: Collagen (F), PCNA expression (G) or transwell invasion (H). Spheroid size and invasion index were normalized to spheroid number. Experimental groups were plated in triplicate. Experiments were repeated three times. Two slides containing three sections each were subject to immunostaining. Mean number of spheroids analyzed/group±s.e.m.: Con, 196±80; HTRA2-KD#2, 219±99; HTRA2-KD#2, 161±40. Statistical analysis was performed using two-tailed t-test (A–D) or one-way ANOVA with Bonferroni post-hoc comparison (E–H). *P<0.05, **P<0.01, ***P<0.001; n.s, not significant. Mean±s.e.m. is shown. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31208996), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human Aldehyde Dehydrogenase 1-A1/ALDH1A1 by Western Blot Effect of p42/44MAPK and SMAD3 knockdown on CCL2/CCR2-mediated ALDH1A1 and HTRA2 expression. (A) DCIS.com, pHAGE or CCR2-H SUM225 cell lines were analyzed for expression of the indicated proteins by immunoblot. Arrow points to SMAD3 protein approximately 50 kda in size. (B) DCIS.com or CCR2-H SUM225 cells were transfected with control siRNAs or siRNAs to p42/44MAPK or SMAD3, stimulated with CCL2 for 24 h and analyzed for expression of the indicated proteins by immunoblot. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31208996), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human Aldehyde Dehydrogenase 1-A1/ALDH1A1 by Western Blot HTRA2 overexpression orknockdown affects CCR2-H SUM225 and CCR2-KO DCIS.com breast cancer cell growth and invasion. (A–D) CCR2-H SUM225 cells expressing vehicle control (pHAGE) or HTRA2 (HTRA2-OE) were analyzed for HTRA2 expression by immunoblot (A), spheroid size in 3D cultures (B), PCNA expression (C) or invasion index (D). Mean number of spheroids analyzed per group±s.e.m.: pHAGE, 269±63; HTRA2-OE, 227±28. (E–H) CCR2-KO DCIS.com cells were transduced with lentivirus expressing Con or HTRA2 shRNAs (HTRA2-KD#2, HTRA2-KD#3) and analyzed for HTRA2 expression by immunoblot (E), spheroid size in Matrigel: Collagen (F), PCNA expression (G) or transwell invasion (H). Spheroid size and invasion index were normalized to spheroid number. Experimental groups were plated in triplicate. Experiments were repeated three times. Two slides containing three sections each were subject to immunostaining. Mean number of spheroids analyzed/group±s.e.m.: Con, 196±80; HTRA2-KD#2, 219±99; HTRA2-KD#2, 161±40. Statistical analysis was performed using two-tailed t-test (A–D) or one-way ANOVA with Bonferroni post-hoc comparison (E–H). *P<0.05, **P<0.01, ***P<0.001; n.s, not significant. Mean±s.e.m. is shown. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31208996), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human Aldehyde Dehydrogenase 1-A1/ALDH1A1 by Western Blot ALDH1A1 knockdown inhibits DCIS.com breast cancer cell growth and invasion. (A–F) DCIS.com cells expressing control shRNAs (Con) or shRNAs to ALDH1A1 (ALDH-KD#1, ALDH-KD#5) were treated with/without 100 ng/ml CCL2 and analyzed for (A) ALDH1A1 and HTRA2 expression by immunoblot, (B) spheroid size in 3D cultures, (C) PCNA expression, (D) transwell invasion, (E) E-cadherin or (F) TWIST1 expression. Mean number of spheroids analyzed per group±s.e.m.: Con, 163+9; Con+CCL2, 165+7; ALDH-KD#1 152+12; ALDH-KD#1+CCL2, 168+11; ALDH-KD#5, 174+10; ALDH-KD#5+CCL2, 162+13. Protein levels in immunoblots were measured by densitometry. Expression levels were normalized to control group. Representative blots of three experiments are shown. Spheroid size and invasion index were normalized to spheroid number. Experimental groups were plated in triplicate. Experiments were performed three times. Two slides containing three sections each were subject to immunostaining. Statistical analysis was performed using one-way ANOVA with Bonferroni post-hoc comparison. *P<0.05, **P<0.01; n.s, not significant. Mean±s.e.m. is shown. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31208996), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human Aldehyde Dehydrogenase 1-A1/ALDH1A1 by Western Blot ALDH1A1 overexpression orknockdown affects CCR2-H SUM225 and CCR2-KO DCIS.com breast cancer cell growth but not invasion. (A–D) CCR2-H SUM225 cells expressing Con shRNAs or shRNAs to ALDH1A1 (ALDH-KD#1, ALDH-KD#5) were analyzed for ALDH1A1 and HTRA2 expression by immunoblot (A), spheroid size in 3D cultures (B), PCNA expression (C) or invasion index (D). Mean number of spheroids analyzed per group±s.e.m.: Con, 197±18; ALDH-KD#1, 190±10; ALDH-KD#5, 196±87. (E–H) CCR2-KO DCIS.com cells expressing vehicle control (pHAGE) or ALDH1A1 (ALDH-OE) were analyzed for: expression of ALDH1A1 and HTRA2 by immunoblot (E), spheroid size in 3D cultures (F), PCNA expression (G) or transwell invasion (H). Protein levels in immunoblots were measured by densitometry. Expression levels were normalized to control group. Representative blots of three experiments are shown. Spheroid size and invasion index were normalized to spheroid number. Experimental groups were plated in triplicate. Experiments were performed three times. Two slides containing three sections each were subject to immunostaining. Mean number of spheroids analyzed per group±s.e.m.: pHAGE, 275±58; ALDH-OE, 219±38. Statistical analysis was performed using one-way ANOVA with Bonferroni post-hoc comparison. *P<0.05; n.s, not significant. Mean±s.e.m. is shown. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31208996), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human Aldehyde Dehydrogenase 1-A1/ALDH1A1 by Western Blot ALDH1A1 overexpression orknockdown affects CCR2-H SUM225 and CCR2-KO DCIS.com breast cancer cell growth but not invasion. (A–D) CCR2-H SUM225 cells expressing Con shRNAs or shRNAs to ALDH1A1 (ALDH-KD#1, ALDH-KD#5) were analyzed for ALDH1A1 and HTRA2 expression by immunoblot (A), spheroid size in 3D cultures (B), PCNA expression (C) or invasion index (D). Mean number of spheroids analyzed per group±s.e.m.: Con, 197±18; ALDH-KD#1, 190±10; ALDH-KD#5, 196±87. (E–H) CCR2-KO DCIS.com cells expressing vehicle control (pHAGE) or ALDH1A1 (ALDH-OE) were analyzed for: expression of ALDH1A1 and HTRA2 by immunoblot (E), spheroid size in 3D cultures (F), PCNA expression (G) or transwell invasion (H). Protein levels in immunoblots were measured by densitometry. Expression levels were normalized to control group. Representative blots of three experiments are shown. Spheroid size and invasion index were normalized to spheroid number. Experimental groups were plated in triplicate. Experiments were performed three times. Two slides containing three sections each were subject to immunostaining. Mean number of spheroids analyzed per group±s.e.m.: pHAGE, 275±58; ALDH-OE, 219±38. Statistical analysis was performed using one-way ANOVA with Bonferroni post-hoc comparison. *P<0.05; n.s, not significant. Mean±s.e.m. is shown. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31208996), licensed under a CC-BY license. Not internally tested by R&D Systems.