PA5-34790

antibody from Invitrogen Antibodies

Targeting: PLAU UPA, URK

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-34790

Provider

Invitrogen Antibodies

Product name

Urokinase Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Recombinant protein fragment

Description

Recommended positive controls: IMR32. Predicted reactivity: Dog (81%), Pig (82%), Rabbit (85%), Sheep (81%). IHC notes, Requires antigen retrieval using heat mediated 10mM Citrate buffer (pH6.0) or Tris-EDTA buffer (pH8.0) Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.

Reactivity

Human

Host

Rabbit

Isotype

IgG

Vial size

100 µL

Concentration

0.77 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot using Urokinase Polyclonal Antibody (Product # PA5-34790). Sample (30 µg of whole cell lysate). Lane A: IMR32. 10% SDS PAGE. Urokinase Polyclonal Antibody (Product # PA5-34790) diluted at 1:2,000.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-Urokinase Polyclonal Antibody (Product # PA5-34790) and a 50 kDa band corresponding to Urokinase was observed in PC-3 and DU 145 but decreased in LNCaP upon treatment with the protein secretory blockers, PTI and FLI-06. Whole cell extracts (30 µg lysate) of PC-3 (Lane 1), PC-3 treated with PTI (1X for 4 hours) (Lane 2), PC-3 treated with FLI-06 (10uM for 4 hours) (Lane 3), DU 145 (Lane 4), DU 145 treated with PTI (1X for 4 hours) (Lane 5), DU 145 treated with FLI-06 (10uM for 4 hours) (Lane 6), LNCaP (Lane 7), LNCaP treated with PTI (1X for 4 hours) (Lane 8) and LNCaP treated with FLI-06 (10uM for 4 hours) (Lane 9) were electrophoresed using NuPAGE™ 10% Bis-Tris Protein Gel (Product # NP0301BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). An uncharacterized band of ~30 kDa was also observed in the positive cell lines panel.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of Urokinase was performed using 70% confluent log phase PC-3 and LNCaP, control and PTI treated, cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with Urokinase Polyclonal Antibody (Product # PA5-34790) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2000 dilution), for 45 minutes at room temperature (Panel a,e,i: Green). Nuclei (Panel b,f,j: Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c,g,k: Blue) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image of untreated PC-3 cells showing faint staining for Urokinase that is enhanced upon PTI treatment (Panel h). Panel l represents LNCaP cells showing no staining for Urokinase upon PTI treatment. The images were captured at 60X magnification.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using Urokinase (Product # PA5-34790) antibody at 1:250 dilution. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.