Invitrogen

tPA Polyclonal Antibody

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tPA Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry analysis of PLAT in paraffin-embedded human placenta tissue. Antigen retrieval was performed on the tissue using citrate buffer (pH 6, 20 min) and blocked with 10% goat serum. Samples were incubated with PLAT polyclonal antibody (Product # PA5-95629) at a 1 µg/mL dilution, followed by biotinylated goat anti-rabbit IgG (30 min, 37°C), and developed with Strepavidin-Biotin-Complex and DAB.

tPA Antibody in Western Blot (WB) — Western blot analysis of PLAT in Lane 1: human HeLa whole cell lysates, Lane 2: human U2OS whole cell lysates, Lane 3: human U-87MG whole cell lysates, Lane 4: rat liver tissue lysates, Lane 5: rat kidney tissue lysates, Lane 6: mouse liver tissue lysates, Lane 7: mouse kidney tissue lysates. Electrophoresis was performed with 5-20% SDS-PAGE gel (70V, Stacking gel; 90V Resolving gel, Time: 2-3 hours), transferred to a nitrocellulose membrane and blocked using 5% Non-fat Milk/TBS (1.5 hrs at room temperature). Samples were incubated with PLAT polyclonal antibody (Product # PA5-95629) using a 0.5 µg/mL dilution, followed by a goat anti-rabbit IgG-HRP at a dilution of 1:10,000, and developed with enhanced chemiluminescence (ECL).

tPA Antibody in Flow Cytometry (Flow) — Flow cytometry of PLAT in U251 cells (blue line), isotype control rabbit IgG (green line) and unlabeled (red line). Samples were blocked with 10% goat serum, incubated with PLAT polyclonal antibody (Product # PA5-95629) at a dilution of 1 µg (per 1x10^6 cells), followed by 488 conjugated goat anti-rabbit IgG (30 min at 20°C) using a 5-10 µg (per 1x10^6 cells) dilution.

Product Details

PA5-95629

Applications	Tested Dilution	Publications
Western Blot (WB)	0.1-0.5 µg/mL	-
Immunohistochemistry (Paraffin) (IHC (P))	0.5-1 µg/mL	-
Immunohistochemistry (Frozen) (IHC (F))	0.5-1 µg/mL	-
Immunocytochemistry (ICC/IF)	0.5-1 µg/mL	-
Flow Cytometry (Flow)	1-3 µg/1x10^6 cells	-

Product Specifications
Species Reactivity	Human, Mouse, Rat
Host/Isotype	Rabbit / IgG
Class	Polyclonal
Type	Antibody
Immunogen	A synthetic peptide corresponding to a sequence of human PLAT. View antigen
Conjugate	Unconjugated
Form	Lyophilized
Concentration	500 µg/mL
Purification	Affinity chromatography
Storage buffer	PBS with 4mg trehalose
Contains	0.05mg sodium azide
Storage conditions	Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
RRID	AB_2807431

Product Specific Information

Synthetic peptide sequence: QKDVPGVYTKVTNYLDWIRDNMRP.

Reconstitution information: Add 0.2 mL of distilled water will yield a concentration of 500 µg/mL.

Target Information

TPA converts the abundant, but inactive, zymogen plasminogen to plasmin by hydrolyzing a single Arg-Val bond in plasminogen. By controlling plasmin-mediated proteolysis, it plays an important role in tissue remodeling and degradation, in cell migration and many other physio pathological events. It specifically cleaves the Arg- -Val bond in plasminogen to form plasmin, and is comprised of a heterodimer of chain A and chain B held by a disulfide bond. TPA binds to fibrin with high affinity. This interaction leads to an increase in the catalytic efficiency of the enzyme between 100-and 1000-fold, due to an increase in affinity for plasminogen. Similarly, binding to heparin increases the activation of plasminogen. Binding to laminin and fibronectin has also been demonstrated. TPA also binds to mannose receptor and the low-density lipoprotein receptor-related protein (LRP1). These proteins are involved in TPA clearance. TPA binds to annexin II and to cytokeratin 8. As yet unidentified interactions on endothelial cells and vascular smooth muscle cells (VSMC) lead to a 100-fold stimulation of plasminogen activation. In addition, binding to VSMC reduces TPA inhibition by PAI-1 by 30-fold.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

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Bioinformatics

Protein Aliases: 3.4.21.68; Alteplase; plasminogen/activator kringle; Reteplase; t plasminogen activator; t-PA; t-plasminogen activator; tissue plasminogen activator; Tissue-type plasminogen activator; Tissue-type plasminogen activator chain A; Tissue-type plasminogen activator chain B

Gene Aliases: AU020998; AW212668; D8Ertd2e; PATISS; PLAT; T-PA; TPA

UniProt ID: (Human) P00750, (Mouse) P11214, (Rat) P19637

Entrez Gene ID: (Human) 5327, (Mouse) 18791, (Rat) 25692

Molecular Function: serine protease