- PA5-11483 - Invitrogen Antibodies ALDH2 antibody

Submitted references Critical role of mitochondrial aldehyde dehydrogenase 2 in acrolein sequestering in rat spinal cord injury.

Herr SA, Shi L, Gianaris T, Jiao Y, Sun S, Race N, Shapiro S, Shi R
Neural regeneration research 2022 Jul;17(7):1505-1511

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of ALDH2 was performed using 70% confluent log phase Hep G2 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with ALDH2 Polyclonal Antibody (Product # PA5-11483) at 1:50 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). Mitochondria (Panel c: red) was stained with Mitotracker Red CMXRos (Product # M7512). Panel d represents the merged image showing mitochondrial localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of A549 cells stained with an ALDH2 polyclonal antibody (Product # PA5-11483). A549 cells were fixed with 4% PFA (20 min), permeabilized with Triton X-100 (0.1%, 10 min), then incubated with an ALDH2 polyclonal antibody (Product # PA5-11483) (1:25, 1 hr at 37°C). Primary antibody was detected with fluor-conjugated donkey anti-rabbit secondary antibody (green) at 1:400 dilution for 50 min at 37°C). Actin filaments have been labeled with dye-conjugated phalloidin (red). Nuclei were counterstained with DAPI (blue) (10 µg/mL, 10 min).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of ALDH2 was performed using 70% confluent log phase Hep G2 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with ALDH2 Polyclonal Antibody (Product # PA5-11483) at 1:50 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). Mitochondria (Panel c: red) was stained with Mitotracker Red CMXRos (Product # M7512). Panel d represents the merged image showing mitochondrial localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunocytochemistry analysis of ALDH2 in A549 cells. Samples were incubated with ALDH2 polyclonal antibody (Product # PA5-11483) using a dilution of 1:25 for 1 h at 37°C followed by Alexa Fluor® 488 conjugated donkey anti-rabbit at a dilution of 1:400 for 50 min at 37°C. Cells were fixed with 4% PFA (20 min) and permeabilized with Triton X-100 (0.1%, 10 min). Cytoplasmic actin was counterstained with Alexa Fluor® 555 (red) conjugated Phalloidin (7 units/mL, 1 h at 37°C). Nuclei were counterstained with DAPI (blue) (10 µg/mL, 10 min). ALDH2 immunoreactivity is localized to Mitochondrion significantly.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 1 Reduction of ALDH2 levels post SCI and its influence by Alda-1 . Acrolein levels were analyzed by western blot (A), with additional confirmation shown by IHC images (B). Groups were divided into ""Control"" for uninjured rats, ""SCI"" for injured rats given vehicle injection, or ""SCI + Alda-1,"" injured rats given 15 mg/kg doses of Alda-1 until sacrifice at 48 hours acute timepoint (total 6 doses). (A) Top: representative ALDH2 bands (50 kDa) are shown above corresponding beta-actin bands (45 kDa). Bottom: Data is normalized to beta-actin and graphed below, showing change as a percent of control. Note the ALDH2 level was suppressed post-SCI (76% of control) which was significantly lower than control ( P < 0.05). However, in the group of ""SCI + Alda-1"", the application of Alda-1 resulted in a trend of recovery of ALDH2 (84% of control) compared with that in SCI. Data are expressed as the mean +- SEM. * P < 0.05 (one-way analysis of variance followed by Tukey's post hoc test). (B) Representative transverse spinal cord sections from each experimental group are immunostained for ALDH2 (brown color). IHC groups are as follows: Control, SCI, and SCI + Alda-1. Note the inconsistent staining after injury. Western blot and IHC: n = 8 and 3-4 for ""Control"", ""SCI"", and ""SCI + Alda-1"" groups, respectively. Alda-1: ALDH2-selective activator; ALDH2: mitochondrial aldehyde dehydrogenase-2; IHC: immunohistochemistry; SCI: spinal cord injury.

PA5-11483

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