Aldolase C-C term, mouse monoclonal, Cat# MCA-1A1

Aldolase C-C term, mouse monoclonal, Cat# MCA-1A1

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View of mixed neuron/glial cultures stained with MCA-1A1 (green) and our rabbit antibody to NeuN/FOX3 antibody (RPCA-FOX3, red). MCA-1A1 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.

Blots of crude rat brain lysates blotted with MCA-1A1. The MCA-1A1 monoclonal binds strongly and cleanly to a band at about 40 kDa. Other studies show that this antibody is completely specific for Aldolase C, and does not react with recombinant human Aldolase A or B.

Above: Blots of recombinant full length His-tagged human Aldolase A, B and C with MCA-1A1 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 human Aldolase A, B and C respectively. The epitope for MCA-1A1 is within the C-terminal peptide of the molecule which is less conserved between the three Aldolases. The specificity of our three Aldolase antibodies is shown in the indicated parts of the image. One antibody, MCA-4A9, binds to the N-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: C-terminus specific.
Reference Code MCA-1A1
HGNC name ALDOC
RRID# AB_2572224
Molecular weight 40 kDa
Immunogen C-terminal sequence KYEGSGEDGGAAAQSLYIANHAY
Isotype IgG1
Concentration Antibody is supplied as 1 mg/mL of affinity purified antibody or concentrated tissue culture supernatant.
Species Reactivity Human, horse, cow, pig, chicken, rat, mouse
Applications Western blot, ICC/IF, IHC
Suggestions for use Western blot: 1:1,000. If/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-1A1 was raised against C-terminal 23 amino acids of aldolase C protein, the sequence is KYEGSGEDGGAAAQSLYIANHAY. 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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