tPA and PAI-1 Deficient Plasma
Affinity’s tPA and PAI-1 Deficient Plasma is manufactured from pooled normal human plasma depleted of both tPA and PAI-1 using antibodies directed to tPA and PAI-1 in separate depletion cycles. Only the highest quality citrated plasma is used as starting material. Our Frozen Deficient or Depleted Plasma products are typically buffered with the addition of HEPES to 20mM final concentration.
All tPA and PAI-1 Deficient Plasma or Depleted Plasma from Affinity Biologicals are for Research Use Only and must not be used for diagnostic applications.
tPA and PAI-1 Deficient – Depleted Plasma
tPA and PAI-1 Deficient – Depleted Plasma is manufactured from normal citrated human plasma that has been depleted of both tPA (Tissue Plasminogen Activator) and PAI-1 (Plasminogen Activator Inhibitor – type 1) by selective affinity immuno-adsorption using antibodies directed towards tPA and PAI-1.
Only the highest quality citrated plasma is used as starting material and in many cases the parent plasma is available as control material. Plasma products are typically buffered with the addition of HEPES to 20mM final concentration and are available in 1ml vials to litre quantities. Our tPA/PAI deficient – depleted plasma can be used for further manufacturing or research use only applications.
Product Code: TPA/PAI-DP
Presentation: Frozen tPA and PAI-1 Deficient – Depleted Plasma
Preparation/Handling: Thaw 1 ml vials in 37oC water bath for 5 minutes; for bulk volumes, thawing time will be dependent on bottle size.
Storage and Stability: Plasma is shipped frozen and should be stored below -60oC. Product is stable until date stated on vial label when stored at -60oC. Once thawed, plasma is stable for 4 hours at 2-8oC in original vial.
Specifications: Plasma is deficient of tPA antigen to ≤0.5 ng/mL and deficient of PAI-1 to <0.5ng/mL – normal for PT, APTT and Fibrinogen levels.
Certificate of Analysis: available upon request
Description of Tissue Plasminogen Activator (tPA)
Tissue-type plasminogen activator (tPA) is one of two major physiologic activators of plasminogen in plasma. It is a serine protease of 68 kDa produced primarily in endothelial cells but is also present in monocytes and megakaryocytes. Normal plasma tPA antigen concentrations have been reported from 20 ng/ml to 5 μg/ml, depending on the assay used, but typically most of the tPA (> 90%) is in complex with it’s primary inhibitor, plasminogen activator inhibitor-1 (PAI-1). Structurally, tPA is a single-chain enzyme that consists of a catalytic domain followed by two kringle structures, an EGF domain and a finger domain. The activation of plasminogen by tPA is dependent on the presence of a fibrin cofactor. The binding of both tPA and plasminogen to fibrin is mediated in part through lysine binding sites within the kringle structures of both enzyme and substrate, but also through the finger domain of tPA. Activation of plasminogen by tPA occurs by cleavage after residue Arg560 to produce the two-chain active serine protease plasmin. The activity of tPA is regulated in part by a very short half life in circulation (t½ of ~4 minutes) and by circulating protease inhibitors PAI-1 and to a lesser extent α2macroglobulin1-3.
References and Reviews
- Bachmann F; The Plasminogen-Plasmin Enzyme System; in Hemostasis and Thrombosis, 3rd Edition, eds. RW Colman, J Hirsh, VJ Marder and EW Salzman, pp. 1592-1622, J.B. Lippincott Co., Philadelphia PA, USA, 1994.
- Holvoet P, Cleemput H, Collen D; Assay of Human Tissue-Type Plasminogen Activator (t-PA) with an Enzyme-Linked Immunosorbent Assay (ELISA) Based on Three Murine Monoclonal Antibodies to t-PA. Thrombosis and Haemostasis 54, pp. 684-687, 1985.
- Giles AR, Nesheim, et al; The fibrinolytic Potential of the Normal Primate following the Generation of Thrombin In Vivo; Thrombosis and Haemostasis 63, pp. 476-481, 1990.
Description of PAI-1 (Type-1 Plasminogen Activator Inhibitor)
Type I plasminogen activator inhibitor, PAI-1, is a 50 kDa single-chain glycoprotein which belongs to the serine protease inhibitor (SERPIN) family of proteins. The concentration of PAI-1 in normal human plasma is very low, with an average concentration of approximately 20 ng/ml. The plasma concentrations of PAI-1 can be affected by a number of factors including diurnal variations, age, sex, pregnancy, obesity and exercise status. PAI-1 is synthesized by various cell types including endothelial cells, hepatocytes, vascular smooth muscle cells, fibroblasts, mesothelial cells, granulosa cells and malignant cell lines. It is also found in the α-granules of platelets as well as plasma. PAI-1 exists in at least three different conformations, including an active form with a half-life of approximately 1 – 3 hours, a latent form and a proteolytically or oxidatively inactivated form. The plasma glycoprotein vitronectin has been shown to bind active PAI-1 and stabilize it in its active conformation, extending its functional half-life to greater than 24 hours. Little specific binding of the latent or inactive forms of PAI-1 to vitronectin occurs. PAI-1 is considered to be the primary regulator of plasminogen activation in vivo. It is the major physiologic inhibitor of both the single chain and two-chain forms of tPA, being able to inhibit the single-chain form at least 10000 times greater than other PAIs.1-3
References and Reviews
- Kruithof, E.K.O. Plasminogen activator inhibitor type 1: biochemical, biological and clinical aspects. Fibrinolysis, 2:59-70, 1988.
- Edelberg, J.M, Sane, D.C., and Pizzo, S.V. Vascular regulation of plasminogen activator inhibitor-1 activity. Semin. Thromb. Hemost., 20(4):319-323, 1994.
- Loskutoff, D.J., Sawdey, M., and Mimuro, J. Type 1 plasminogen activator inhibitor, Prog. Thromb. Haemost., 9:87-115, 1988.
TPA/PAI-DP – Tissue Plasminogen Activator (tPA) and Plasminogen Activator Inhibitor – Type 1 (PAI-1) Deficient Plasma – 1ml or bulk quantities
TPA-DP – Tissue Plasminogen Activator (tPA) Deficient Plasma – 1ml or bulk quantities
TPA-EIA – Tissue Plasminogen Activator – tPA Paired Antibody Set – 5 plate set
SATPA-AP – Sheep anti human tPA – Tissue-type Plasminogen Activator, affinity purified IgG (0.5 mg vial)
SATPA-HRP – Sheep anti human tPA – Tissue-type Plasminogen Activator, peroxidase conjugated IgG (0.2 mg vial)
PAI-DP – Frozen PAI-1 Deficient Plasma – 1ml or bulk quantities
PAI-LDP – Lyophilized PAI-1 Deficient Plasma – 1ml
SAPAI-HRP – Sheep anti human PAI-1 – Plasminogen Activator Inhibitor type 1, peroxidase conjugated IgG (0.2 mg vial)