Aldolase C-N term, mouse monoclonal, Cat# MCA-4A9

Aldolase C-N term, mouse monoclonal, Cat# MCA-4A9

Left: View of mixed neuron/glial cultures stained with MCA-4A9 (green) and our rabbit antibody to NeuN/FOX3 antibody (RPCA-FOX3, red). MCA-4A9 antibody reveals strong cytoplasmic staining in astrocytes, while Rabbit Fox3/NeuN antibody shows nuclear and distal cytoplasmic staining in neuron cells and is complete absence of astrocytes. Blue is a DNA stain. Middle and Right: Mouse brain sections (fixed by transcardial perfusion with 4% paraformaldehyde) stained with MCA-4A9 (red) and our chicken anti-Vimentin CPCA-Vim antibody (green). In the striatum (Middle), MCA-4A9 positive astrocytes are highly co-stained with CPCA-Vim, which results in yellow to gold colors. In the cerebellum (Right), however, MCA-4A9 positive Purkinje cells do not express vimentin, which results in red color. Insets show a higher magnification picture of MCA-4A9 single labeling in red. Nuclei are labeled with Dapi (blue).

Blots of crude cow cerebellum homogenate blotted with MCA-4A9. The MCA-4A9 monoclonal binds strongly and cleanly to a band at about 40 kDa. Other studies show that this antibody does not recognize the very closely related molecules aldolase A or B.

Above: Blots of recombinant human Aldolase A, B and C with MCA-4A9 as indicated showing binding to only the Aldolase C gene product, with no cross reactivity to the closely related Aldolases A and B. 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-4A9 is within the N-terminal peptide of the molecule which is less conserved between the three Aldolases. The specificity of our other two Aldolase antibodies is shown in the indicated parts of the image. One antibody, MCA-1A1, binds to the C-terminal peptide and also is specific for Aldolase C, while another, MCA-E9, binds to the less conserved core of the molecule and recognizes all three Aldolase C gene products.

Product name Anti-Aldolase C
Description Mouse Monoclonal to Anti-Aldolase C: N-terminus specific.
Reference Code MCA-4A9
HGNC name ALDOC
RRID# AB_2571880
Molecular weight 40 kDa
Immunogen N-terminal sequence MPHSYPALSAEQKKELSDIA
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: 1:1,000. IF/ICC or IHC: 1:500-1:1,000.
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-4A9 was raised against N-terminal 20 amino acids of aldolase C protein, the sequence MPHSYPALSAEQKKELSDIA, so that the epitope is unusually well known. 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).

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