Journal:

Article Title: Retinoid Metabolism and ALDH1A2 (RALDH2) Expression are Altered in the Transgenic Adenocarcinoma Mouse Prostate Model

doi: 10.1016/j.bcp.2009.06.022

Figure Lengend Snippet: Primer Sequences Used for Quantitative RT-PCR

Article Snippet: 2.5 ALDH1A2 Polyclonal Antibody Generation A rabbit anti-mouse ALDH1A2 polyclonal antibody was generated by contract with Alpha Diagnostics International (San Antonio, TX).

Techniques: Sequencing

Journal:

Article Title: Retinoid Metabolism and ALDH1A2 (RALDH2) Expression are Altered in the Transgenic Adenocarcinoma Mouse Prostate Model

doi: 10.1016/j.bcp.2009.06.022

Figure Lengend Snippet: A, Total RNA was extracted and reverse-transcribed from prostate lobes microdissected from three nontransgenic mice at 18, 24, and 36 weeks of age. The bars indicate the average transcript levels of ALDH1A1, ALDH1A2, and ALDH1A3 in prostate tissue from age-matched nontransgenic controls (WT, black bars) and TRAMP (white bars) normalized to 36B4 mRNA levels in each sample. B, Relative mRNA levels of RARβ2, CYP26A1, and LRAT measured by quantitative RT-PCR in WT (black bars) and TRAMP mice (white bars) at 36 weeks of age. All samples were normalized to 36B4. VP, ventral prostate; LP, lateral prostate; DP, dorsal prostate; and AP, anterior prostate. Relative expression was calculated using the Bio-Rad Genex software, where all prostate lobes from a given age group were processed independently. Error bars= standard error. Comparisons for statistical analysis were made for each lobe between the Wt and the TRAMP mice at each of the time points. *, p ≤ 0.05 as determined by two-tailed Student's t test, comparing TRAMP mice to their age-matched littermate controls. (TRAMP = TRAMP+).

Article Snippet: 2.5 ALDH1A2 Polyclonal Antibody Generation A rabbit anti-mouse ALDH1A2 polyclonal antibody was generated by contract with Alpha Diagnostics International (San Antonio, TX).

Techniques: Reverse Transcription, Quantitative RT-PCR, Expressing, Software, Two Tailed Test

Journal:

Article Title: Retinoid Metabolism and ALDH1A2 (RALDH2) Expression are Altered in the Transgenic Adenocarcinoma Mouse Prostate Model

doi: 10.1016/j.bcp.2009.06.022

Figure Lengend Snippet: A, Wild type mouse embryo at E12/13 (WT). ALDH1A2 staining located in the metanephros. B, ALDH1A2-/- embryo at E12/13. No ALDH1A2 staining was observed in the metanephros, confirming antibody specificity. E, Wild type mouse testis at 36 weeks of age. ALDH1A2 staining located in the germ cells but not in the spermatagonia. G, Wild type mouse kidney at 36 weeks of age. ALDH1A2 staining was observed in the tubular cells, but not in the glomeruli. C-D and G-H, Negative control incubated with preimmune serum instead of primary antibody. A-D, 200× magnification. E-H, 400× magnification.

Article Snippet: 2.5 ALDH1A2 Polyclonal Antibody Generation A rabbit anti-mouse ALDH1A2 polyclonal antibody was generated by contract with Alpha Diagnostics International (San Antonio, TX).

Techniques: Staining, Negative Control, Incubation

Journal:

Article Title: Retinoid Metabolism and ALDH1A2 (RALDH2) Expression are Altered in the Transgenic Adenocarcinoma Mouse Prostate Model

doi: 10.1016/j.bcp.2009.06.022

Figure Lengend Snippet: Tissue extracts were obtained from microdissected dorsal prostate lobes (30 μg protein loaded/lane) from nontransgenic (labeled as WT) littermate controls and TRAMP mice at 18, 24, and 36 weeks of age (3 mice per condition). A, Western blot analysis performed with the polyclonal rabbit anti-mouse ALDH1A2 antibody. B, Western blot analysis performed with the polyclonal rabbit anti-human S100A4 antibody. For a loading control, these blots were stripped and reblotted with a polyclonal goat anti-human actin antibody. Positive controls: ALDH1A2-wild type testis extract (15 μg protein loaded/lane); S100A4-PC3 cell extract (30 μg protein loaded/lane). This experiment was performed three times with similar results; one blot is shown. The upper arrow at the right in panel A shows ALDH1A2; the lower arrow points to the non-specific protein band at ∼30 kd. C, Densitometric quantitation of similar ALDH1A2 and S100A4 Western Blots. Band density was measured using the Fluor Chem 8800 software (Alpha Innotech) for the bands from ALDH1A2 (left) and S100A4 (right). Western blots normalized to actin for all prostate samples. Error bars = standard error. *, p ≤ 0.05 as determined by two-tailed Student's t test. TRAMP mice were compared to age-matched littermate nontransgenic controls (WT). DP, dorsal prostate.

Article Snippet: 2.5 ALDH1A2 Polyclonal Antibody Generation A rabbit anti-mouse ALDH1A2 polyclonal antibody was generated by contract with Alpha Diagnostics International (San Antonio, TX).

Techniques: Labeling, Western Blot, Control, Quantitation Assay, Software, Two Tailed Test

Journal:

Article Title: Retinoid Metabolism and ALDH1A2 (RALDH2) Expression are Altered in the Transgenic Adenocarcinoma Mouse Prostate Model

doi: 10.1016/j.bcp.2009.06.022

Figure Lengend Snippet: All tissue sections were stained with hematoxylin (blue). A, C, E, and G, ALDH1A2 staining. B, D, F, and H, S100A4 staining. A-H, 600× magnification. Dorsal lobe (A) and lateral lobe (C) from an 18 week old nontransgenic mouse. Strong nuclear and cytoplasmic ALDH1A2 staining (brown stain) representative of normal prostate epithelial cells in all lobes. S100A4 (brown stain) staining in fibroblast cells surrounding a normal prostate gland in dorsal lobe (B) and lateral lobe (D). E, dorsal prostate tumor tissue in a 36 week old TRAMP mouse, showing weak cytoplasmic ALDH1A2 staining in prostate cancer cells with little to no stain in the nuclei. There is no ALDH1A2 staining in the stroma of dorsal prostate tissue in the TRAMP mouse. F, S100A4 staining in the stroma of TRAMP mice and among prostate cancer cells within the gland. G, lateral prostate tumor tissue in a 36 week old TRAMP mouse. The ALDH1A2 staining pattern in the prostate cancer cells is similar to E. Positive staining of ALDH1A2 in the lateral prostate stroma, which is unique to this lobe. F, S100A4 staining in the prostate cancer and lateral prostate stroma. I-J, Negative controls incubated with preimmune serum instead of primary antibody. I, prostate tumor tissue from dorsal prostate of 30 week old TRAMP mouse, 200× magnification. J, prostate tumor tissue from lateral prostate of 30 week old TRAMP mouse, 200× magnification. K, ALDH1A2 staining of prostate tumor tissue from 30 week old TRAMP mouse, 200× magnification. L, adjacent section of ALDH1A2 staining of prostate tumor tissue from 30 week old TRAMP mouse. Tumor tissue section was simultaneously incubated with the 20× peptides to which the antibody was generated. (Arrows indicate areas of sections discussed in the Results section).

Article Snippet: 2.5 ALDH1A2 Polyclonal Antibody Generation A rabbit anti-mouse ALDH1A2 polyclonal antibody was generated by contract with Alpha Diagnostics International (San Antonio, TX).

Techniques: Staining, Incubation, Generated

Journal:

Article Title: Retinoid Metabolism and ALDH1A2 (RALDH2) Expression are Altered in the Transgenic Adenocarcinoma Mouse Prostate Model

doi: 10.1016/j.bcp.2009.06.022

Figure Lengend Snippet: A-J, representative slides of a human prostate tumor tissue. A-B and E-F, ALDH1A2 staining at 200× and 600× (boxed areas of A and E) magnification. B and F, ALDH1A2 staining located in the cytoplasmic compartment of basal and luminal cells in a normal prostate gland with weak staining in adjacent cancer cells. C-D and G-H, S100A4 staining at 200× and 600× (boxed areas of C and G) magnification. S100A4 staining is seen in benign glands, as well as in surrounding stroma. I-J, Negative controls, prostate specimens incubated with preimmune serum instead of primary antibody, 200× magnification. (Arrows indicate areas on sections discussed in the Results section).

Article Snippet: 2.5 ALDH1A2 Polyclonal Antibody Generation A rabbit anti-mouse ALDH1A2 polyclonal antibody was generated by contract with Alpha Diagnostics International (San Antonio, TX).

Techniques: Staining, Incubation

Journal:

Article Title: Alcohol-Induced Oxidative Stress in the Liver

doi: 10.1007/978-1-59745-242-7_14

Figure Lengend Snippet: Effect of LPS on 4OH-nonenal adduct accumulation in mouse liver as determined by quantitative immunohistochemistry. Representative photomicrographs are shown depicting 4OH-nonenal immunostaining of a liver section (100×) of a mouse 24 h after injection with LPS (10 mg/kg intraperitoneally). The same microscope region is shown before (A) and after (B) white balancing of the image-analysis software. Note that the acellular space in (A) is light yellow instead of white. (C) is the same as (B), but after the blue exposure is increased 10%. Note that the contrast between the DAB chromogen (brown) and the hematoxylin counterstain (blue) is enhanced by digitally increasing the blue exposure. (D) shows the microscope region from (C), determined by the computer to be within the threshold range for positive staining

Article Snippet: Rabbit polyclonal anti-4OH-nonenal antibody (Alpha Diagnostic); Rabbit polyclonal anti-nitrotyrosine antibody (Upstate).

Techniques: Immunohistochemistry, Immunostaining, Injection, Microscopy, Software, Staining