Platelet Derived Growth Factor AA,AB,BB Neutralizing (PDGF-AA,AB,BB)

Platelet-Derived Growth Factor (PDGF) is the major serum mitogen for cells of mesenchymal origin in humans. The biologically active protein is a dimer composed of two related polypeptides designated A and B. The PDGF protein has been implicated both directly as well as indirectly in several pathological states including neoplasia, arthritis, arteriosclerosis and bone marrow sclerosis. \n\nApplications:\nSuitable for use in Western Blot and Growth Inhibition Assays. Other applications have not been tested.\n\nRecommended Dilution:\nWestern Blot: 5-10ug/ml, under non-reducing conditions, detects 100ng of human PDGF-AB (MW~28kD) as well as 100ng of human PDGF-BB (MW~30-32kD). \nGrowth Inhibition Assay: In a 5 day assay, 10-20ug/ml inhibited 3T3 cell growth by 100% as stimulated by 10ng/ml of PDGF-AB. \nOpimal dilutions to be determined by the researcher.\n\nGrowth Inhibition Assay Protocol:\n1. In DMEM, 1% calf serum, preincubate PDGF-AB (10ng/ml) and P4258-19, PDGF (10-20ug/ml) for one hour at 37 degrees C. Serially dilute P4258-19 2-fold from 40ug/ml to 2.5ug/ml in a 96 well flat black view plate.\n2. Prepare a 3T3 cell suspension at 10e4 cells/ml and aliquot 100ul to each well.\n3. Refeed on Day 3.\n4. Count wells on Day 5 using an ATP endpoint assay. Wash cell layer once with PBS prior to lysis.\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20 degrees C. Aliquots are stable for 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

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SPECIFICATIONS

Catalog Number

P4258-19

Size

1mg

Applications

WB

Hosts

Goat

Reactivities

Hum

Form

Supplied as a liquid in 0.1M Tris-glycine, pH 7.0.

P Type

Pab

Purity

Purified by DEAE chromatography.

Isotype

IgG

References

1. Feakins, R. M., et al. Hum Pathol, 31:1214-22 (2000). Platelet-derived growth factor expression in phyllodes tumors and fibroadenomas of the breast. 2. Gorlach, A., et al. Faseb J, 14:1518-28 (2000). Oxidative stress and expression of p22phox are involved in the up-regulation of tissue factor in vascular smooth muscle cells in response to activated platelets. 3. Shimokawa, H., et al. J Clin Invest, 97:769-76 (1996). Chronic treatment with interleukin-1 beta induces coronary intimal lesions and vasospastic responses in pigs in vivo. The role of platelet-derived growth factor. 4. Ko, F. N., et al. J Clin Invest, 98:1493-501 (1996). Coagulation factor Xa stimulates platelet-derived growth factor release and mitogenesis in cultured vascular smooth muscle cells of rat. 5. Williams, Science 243:1564.

Additional Info

Recognizes non-reduced human PDGF AA, AB and BB dimers in Western Blot. Species Crossreactivity: porcine and bovine PDGF may be detected with decreased sensitivity. Rodent PDGF will not be detected. This antibody will neutralize human, porcine, bovine and rodent PDGF.