The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/1000 - 1/10000. Detects a band of approximately 62 kDa (predicted molecular weight: 62 kDa).
1/100 - 1/250.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
Irreversible conversion of delta-1-pyrroline-5-carboxylate (P5C), derived either from proline or ornithine, to glutamate. This is a necessary step in the pathway interconnecting the urea and tricarboxylic acid cycles. The preferred substrate is glutamic gamma-semialdehyde, other substrates include succinic, glutaric and adipic semialdehydes.
Highest expression is found in liver followed by skeletal muscle, kidney, heart, brain, placenta, lung and pancreas.
Amino-acid degradation; L-proline degradation into L-glutamate; L-glutamate from L-proline: step 2/2.
Involvement in disease
Defects in ALDH4A1 are the cause of hyperprolinemia type 2 (HP-2) [MIM:239510]. HP-2 is characterized by the accumulation of delta-1-pyrroline-5-carboxylate (P5C) and proline. The disorder may be causally related to neurologic manifestations, including seizures and mental retardation.
Immunofluorescent analysis of -20℃ acetone fixed HeLa cells labeling ALDH4A1 with ab185208 at 1/250 dilution (red). Secondary antibody: Goat anti rabbit IgG (Alexa Fluor®555) at a 1/200 dilution. Counterstained with Dapi (blue).
Flow cytometric analysis of 2% paraformaldehyde fixed HepG2 cells labeling ALDH4A1 with ab185208 at 1/160 dilution (red), or a rabbit IgG isotype control (green), followed by secondary goat anti rabbit IgG (FITC) at 1/150 dilution.
Western blot - Anti-ALDH4A1 antibody [EPR14287] (ab185208)
Anti-ALDH4A1 antibody [EPR14287] (ab185208) at 1/5000 dilution + HepG2 cell lysate at 20 µg
Secondary Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution