Aldolase A, B and C mouse monoclonal, Cat# MCA-E9

Aldolase A, B and C mouse monoclonal, Cat# MCA-E9

View of mixed neuron/glial cultures stained with MCA-E9 (red) and our chicken antibody to GFAP antibody (CPCA-GFAP, green). MCA-E9 antibody reveals strong cytoplasmic staining in astrocytes. Blue is a DNA stain. Aldolase stains the astrocytes cell body and processes, whereas GFAP labels the intermediate filament of the cytoskeleton in subset of astrocytes.

Right: Blots of rat brain lysate blotted with MCA-E9. The MCA-E9 monoclonal binds strongly and cleanly to a band at about 40 kDa. We now know that this antibody binds to an epitope shared by the closely related Aldolase A, B and C, recognizing all three on western blots.

Above: Blots of recombinant human Aldolase A, B and C with MCA-E9 as indicated showing binding to all three gene products. Lane labelled S show molecular weight standards, while lanes A, B and C contain recombinant full length His-tagged human Aldolase A, B and C respectively. The epitope for MCA-E9 is within the core of the three Aldolase molecules which are less conserved than the N and C-termini where our Aldolase C specific antibodies bind. The specificity of our other two Aldolase antibodies is shown in the indicated parts of the image. One antibody binds to the N-terminal peptide MCA-4A9 and one to the C-terminal peptide MCA-1A1 bind.

Product name Anti-Aldolase C
Description Mouse Monoclonal to Aldolase C
Reference Code MCA-E9
HGNC name ALDOC
RRID# AB_2572223
Molecular weight 40 kDa
Immunogen Recombinant human full-length aldolase C
Isotype IgG1
Concentration Antibody is supplied as an aliquot of 1 mg/mL of affinity purified antibody.
Species Reactivity Human, horse, cow, pig, chicken, rat, mouse
Applications Western blot, ICC/IF, IHC
Suggestions for use Western blot: 1:10,000. IF/ICC or IHC: 1:1,000-1:5,000.
Storage instructions Shipped on ice. Store at 4°C. For long term storage, leave frozen at -20°C. Avoid freeze / thaw cycles.


Aldolases are glycolytic enzymes that catalyze the reversible aldol cleavage of fructose 1,6-bisphosphate and fructose-1-phosphate to dihydroxyacetone phosphate and either glyceraldehyde 3-phosphate or glyceraldehyde, respectively. Three aldolase isozymes are found in mammals specifically aldolases A, B, and C, each of which is encoded by a separate gene.

Aldolase A is generally considered to be a muscle enzyme. Northern analysis of cultured cells suggests that it is present in both neurons and glia (1). Aldolase B is considered to be a liver-specific enzyme and it is transcriptionally activated by signals from hormones and dietary factors (2). In the adult, aldolase C is the brain-specific isozyme, with low but detectable activity in fetal tissues (1, 3-6). Aldolase C shares 81% amino acid identity with aldolase A and 70% identity with aldolase B.

Earlier studies using isozyme-specific antibodies report its location in gray matter astrocytes and cells of the pia mater (5, 8). In situ hybridization of mouse central nervous system using isozyme-specific probes revealed that aldolase A and C are expressed in complementary cell types: aldolase A mRNA is found in neurons; aldolase C message is detected in astrocytes, some cells of the pia mater, and Purkinje cells (9). Aldolase C can in some situations be used as an astrocyte marker. However Purkinje cells of the cerebellum contain high levels of the enzyme, so the enzyme is not totally astrocyte specific.

MCA-E9 was t raised against full length recombinant human Aldolase C protein expressed and purified from E. coli. The HGNC name for this protein is ALDOC.


References:

1. Popovici T, Berwald-Netter Y, Vibert M, Kahn A, Skala H. Localization of aldolase C mRNA in brain cells. FEBS Lett. 268, 189-193 (1990).

2. Weber A, Marie J, Cottereau D, Simon M, Besmond C, Dreyfus J. & Kahn A. Dietary Control of Aldolase B and L-type Pyruvate Kinamse RNAs in Rat.  J. Biol. Chem 259, 1798-1802 (1984).

3. Mukai T, Yatsuki H, Masuko S, Arai Y, Joh K & Hori K. The structure of the brain-specific rat aldolase C gene and its regional expression. Biochem. Biophys. Res. Commun. 174, 1035-1042 (1991).

4. Royds J, Ironside J, Warnaar S, Taylor C & Timperle W. Monoclonal antibody to aldolase C: a selective marker for Purkinje cells in the human cerebellum. Neuropathol. Appl. Neurobiol. 13, 11-21(1987).

5. Thompson R., Kynoch P. Willson V. Cellular localization of aldolase C subunits in human brain. Brain Res. 232, 489-493 (1982).

6. Schapira F, Reuber M, Hatzfeld A. Resurgence of two fetal-type of aldolases (A and C) in some fast-growing hepatomas. Biochem. Biophys. Res. Commun. 40, 321-327(1970).

7. Arai Y, Kajihara S, Masuda J, Ohishi S, Zen K, Ogata J. Mukai T. Position-independent, high-level, and correct regional expression of the rat aldolase C gene in the central nervous system of transgenic mice. Eur. J. Biochem. 221, 253-260 (1994).

8. Wachsmuth E, Thorner M. & Pfleiderer G. The cellular distribution of aldolase isozymes in rat kidney and brain determined in tissue sections by the immuno-histochemical method. Histochemistry, 45, 143-161 (1975).

9. Walther EU, Dichgans M, Maricich SM, Romito RR, Yang F, Dziennis S, Zackson S, Hawkes R, Herrup K. Genomic sequences of aldolase C (Zebrin II) direct lacZ expression exclusively in non-neuronal cells of transgenic mice. Proc Natl Acad Sci U S A. Mar 3;95(5):2615-20(1998).

Download Adobe Acrobat format product data sheet: .