Product nameAnti-ASPA antibody
See all ASPA primary antibodies
DescriptionRabbit polyclonal to ASPA
Tested applicationsSuitable for: ICC/IF, WB, IHC-P, IPmore details
Species reactivityReacts with: Human
Predicted to work with: Mouse, Cow, Pig
Recombinant fragment, corresponding to a region within amino acids 39-270 of Human ASPA (AAH29128).
- 293T, A431, H1299, or HeLa cell lines. MOLT4 and Raji whole cell lysate. Cal27 xenograft IF/ICC: U-87 MG cell line.
Storage instructionsShipped at 4°C. Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage bufferpH: 7.00
Preservative: 0.01% Thimerosal (merthiolate)
Constituents: 1.21% Tris, 0.75% Glycine, 20% Glycerol
Concentration information loading...
PurityImmunogen affinity purified
- Pathways and Processes
- Metabolic signaling pathways
- Energy transfer pathways
- Energy Metabolism
Our Abpromise guarantee covers the use of ab97454 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use a concentration of 5 µg/ml.|
|WB||1/500 - 1/3000. Predicted molecular weight: 36 kDa.|
|IHC-P||1/100 - 1/500.|
|IP||Use a concentration of 5 µg/ml.|
FunctionCatalyzes the deacetylation of N-acetylaspartic acid (NAA) to produce acetate and L-aspartate. NAA occurs in high concentration in brain and its hydrolysis NAA plays a significant part in the maintenance of intact white matter. In other tissues it act as a scavenger of NAA from body fluids.
Tissue specificityBrain white matter, skeletal muscle, kidney, adrenal glands, lung and liver.
Involvement in diseaseDefects in ASPA are the cause of Canavan disease (CAND) [MIM:271900]; also known as spongy degeneration of the brain. CAND is a rare neurodegenerative condition of infancy or childhood characterized by white matter vacuolization and demeylination that gives rise to a spongy appearance. The clinical features are onset in early infancy, atonia of neck muscles, hypotonia, hyperextension of legs and flexion of arms, blindness, severe mental defect, megalocephaly, and death by 18 months on the average.
Sequence similaritiesBelongs to the AspA/AstE family. Aspartoacylase subfamily.
Cellular localizationCytoplasm. Nucleus.
- Information by UniProt
- ACY 2 antibody
- ACY-2 antibody
- ACY2 antibody
All lanes : Anti-ASPA antibody (ab97454) at 1/1000 dilution
Lane 1 : MOLT4 whole cell lysate
Lane 2 : Raji whole cell lysate
Lysates/proteins at 30 µg per lane.
Predicted band size: 36 kDa
10% SDS PAGE
ab97454 stained U-87 MG cells. The cells were 100% methanol fixed for 5 minutes at -20°C and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1hour at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab97454 at 5µg/ml) overnight at +4°C. The secondary antibody (pseudo-colored green) was Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) used at a 1/1000 dilution for 1hour at room temperature. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.
ASPA was immunoprecipitated using 0.5mg MOLT4 whole cell extract, 5µg of Rabbit polyclonal to ASPA and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, MOLT4 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab97454.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 36kDa, non specific bands - 42 and 51kDa: We are unsure as to the identity of this extra band; ASPA
ab97454 at 1/100 dilution staining ASPA in paraffin-embedded Cal27 xenograft by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).