ALEXIS Biochemicals: Monoclonal Antibody to Matrix Gla Protein (undercarboxylated) (BD4.4)

ALX-804-642

Revised 22-Apr-08

Monoclonal Antibody to Matrix Gla Protein (undercarboxylated) (BD4.4)
SYNONYMS	anti-ucMGP MAb (BD4.4) anti-GluMGP MAb (BD4.4)
PRODUCT LINE	Bone Metabolism
PRODUCT CATEGORY	Osteocalcin & Matrix Gla-protein [MGP] Family / Related Products

Ordering Information

Product Numbers:	Format:	Size:	Unit Price:	Quantity:	Add To Cart
ALX-804-642-C100		100 µg	490.00 USD

Product Specification

SPECIES CROSSREACTIVITY:	Human Rat Others
CLONE:	BD4.4
ISOTYPE:	Mouse IgG1
CONCENTRATION:	0.7mg/ml after reconstitution.
PURITY DETAIL:	Protein G-affinity purified.
FORMULATION:	Lyophilized from PBS containing 0.05% sodium azide.
RECONSTITUTION:	Reconstitute with 145µl distilled water.
IMMUNOGEN:	Synthetic peptide corresponding to the non-carboxylated domain (aa 35-49) of human MGP (matrix Gla protein).
SPECIFICITY:	Recognizes human and rat non-carboxylated (non-functional) MGP.
APPLICATION:	ELISA (1:1’000) Immunohistochemistry (frozen sections (1:250), paraffin sections (1:250)) Western Blot (1:10’000)
SHIPPING:	SHIPPED ON BLUE ICE
SHORT TERM STORAGE:	+4°C
LONG TERM STORAGE:	-20°C
USE/STABILITY:	Stable for at least 1 month when stored at +4°C.
HANDLING:	Avoid freeze/thaw cycles. After opening, prepare aliquots and store at -20°C.

Product Images

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Product Specific Literature References

Novel conformation-specific antibodies against matrix gamma-carboxyglutamic acid (Gla) protein: undercarboxylated matrix Gla protein as marker for vascular calcification: L.J. Schurgers, et al.; Arterioscler. Thromb. Vasc. Biol. 25, 1629 (2005) Abstract; Full Text

General Information

Matrix Gla protein (MGP) is a potent inhibitor of vascular calcification. MGP is mainly synthesized and expressed in cartilage and the vessel wall, where it is produced by the chondrocytes and vascular smooth muscle cells, respectively. During biosynthesis, MGP undergoes two forms of post-translational modifications: the phosphorylation of 3 serine residues and the γ-carboxylation of 5 glutamate residues. Phosphorylation of MGP is essential for its regular cellular secretion, and the vitamin K-dependent carboxylation is crucial for the calcification inhibitory activity. MGP plays a role in bone metabolism and atherosclerosis.

BACKGROUND/TECHNICAL INFORMATION

Immunohistochemistry Protocol
Staining of paraffin embedded tissue sections fixed with 4.5% neutrally buffered formalin:
1. Deparaffinize and rehydrate tissue sections (100% xylol 2x; 100% ethanol; 96% ethanol;
    70% ethanol; 50% ethanol).
2. Wash sections in TBST.
3. Heat 0.2% citric acid, pH 6.0, in microwave until boiling.
4. Put sections in pre-warmed citric acid and "keep-warm" in the microwave for 15 min.
5. Cool down sections for 15 min. and wash three times in TBST.
6. Apply MAb to Matrix Gla Protein (undercarboxylated) (BD4.4) (Prod. No. ALX-804-642) (1:250) in TBST containing 1% BSA and incubate overnight at +4°C.
7. Wash sections 3x in TBST.
8. Use appropriate ABC detection kit (e.g. VECTASTAIN ABC Kit) according to manufacturer’s
    instructions.
9. Apply and stain sections with substrate.
10. Wash sections in tap water for 10 min.
11. Counterstain sections with haemalaun (20 sec.).
12. Wash sections in tap water for 10 min.
13. Coverslip section using an aqueous mounting media.

Western Blot Recommendations
1. Block membrane with PBST containing 3% BSA for at least 1 hour at room temperature.
2. Wash 3x in PBST.
3. Incubate overnight at +4°C with MAb to Matrix Gla Protein (undercarboxylated) (BD4.4) (Prod. No.
    ALX-804-642) (1:10’000) in PBST containing 1% BSA.

General Literature References

The cDNA and derived amino acid sequences for human and bovine matrix Gla protein: M.C. Kiefer, et al.; Nucleic Acids Res. 16, 5213 (1988) Abstract; Full Text
Molecular structure, chromosome assignment, and promoter organization of the human matrix Gla protein gene: L. Cancela, et al.; J. Biol. Chem. 265, 15040 (1990) Abstract; Full Text
Post-translational modifications regulate matrix Gla protein function: importance for inhibition of vascular smooth muscle cell calcification: L.J. Schurgers, et al.; J. Thromb. Haemost. 5, 2503 (2007) Abstract

ALX-804-643	Monoclonal Antibody to Matrix Gla Protein (carboxylated) (G8A.1)
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