Clone #

:

3C

Antigen

:

MG-modified keyhole-lympet hemocyanine

Specificity

:

Specific for MG-modified protein (especially Argpyrimidine)

Subclass

:

Mouse IgG2a

Form

Protein A purified.
Frozen (10mM PBS containing 0.1 %NaN3 and 0.5% BSA).
100 micro g/mL IgG.

Application

Immunohistochemistry?It is recommended that the antibody
be tried at 0.5-1.0 micro g/mL on paraformaldehyde fixed tissue

Storage

Less than -20°C

Paraffin sections were deparaffinized and rehydrated.

Sections were quenched for several minutes with 3% hydrogen
peroxide, rinsed in PBS, pretreated for 30 min with 3% nonimmune
animal serum in PBS.

Antigen retrieval by microwave treatment.

Sections were incubated overnight at 4°C with a antibody
at a concentration of 0.5-10 micro gram/mL.

Antibody binding was visualized by the avidin-biotin-immuno peroxidase
complex method using the Vectastain ABC kit (Vector, Burlingame, CA)
according to the manufacturer's instructions.

3,3-Diaminobenzidine tetrahydrochloride was used as the chromogen.

Hematoxylin or Eosin was used as the counter stain.

Sections from which the primary antibodies were omitted served
as negative reaction controls.

For frozen materials, samples were fixed in 10% formalin, immersed
in 30% sucrose in PBS, embedded at OCT (Sakura, Tokyo, Japan),
and stored at 80°C.

For paraffin embedded materials, samples were fixed in 10% formalin, dehydrated, embedded in paraffin, and stored at room temperature.

Microwave treatment improves the target staining, whereas it declines
non-specific background.

Protease K treatment declines the target staining.

1)

Protein modification by a Maillard reaction intermediate methylglyoxal. Immunochemical detection of fluorescent 5-methylimidazolone derivatives in vivo.
Uchida K, Khor OT, Oya T, Osawa T, Yasuda Y, Miyata T
FEBS Lett. 410(2-3), p313-318. (1997)

2)

Methylglyoxal Modification of Protein -Chemical and Immunochemical Characterization of Methylglyoxal-Arginine Adduct.
T.Oya, N.Hattori, Y.Mizuno, S.Miyata, S.Maeda, T.Osawa and K.Uchida
J.Biological Chemistry, 274, 18492-18502 (1999)

3)

Advanced glycation and lipidoxidation of the peritoneal membrane: respective roles of serum and peritoneal fluid reactive carbonyl compounds.
Miyata T, Horie K, Ueda Y, Fujita Y, Izuhara Y, Hirano H, Uchida K, Saito A, van Ypersele de Strihou C, Kurokawa K
Kidney Int. 58(1), p425-435.(2000)

4)

Argpyrimidine, a blue fluorophore in human lens proteins: high levels in brunescent cataractous lenses.
Padayatti PS, Ng AS, Uchida K, Glomb MA, Nagaraj RH
Invest Ophthalmol Vis Sci. 42(6), p1299-1304. (2001)

5)

High concentrations of glucose induce synthesis of argpyrimidine in retinal endothelial cells.
Padayatti PS, Jiang C, Glomb MA, Uchida K, Nagaraj RH
Curr Eye Res. 23(2), p106-115. (2001)

6)

Curcumin Inhibits ROS Formation and Apoptosis in Methylglyoxal- Treated Human Hepatoma G2 Cells.
WEN-HSIUNG CHAN, HSIN-JUNG WU and YAN-DER HSUUW
Ann. N.Y. Acad. Sci. 1042: 372-378 (2005)

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