Overview

  • Product name
  • Description
    Rabbit polyclonal to Urokinase
  • Host species
    Rabbit
  • Specificity
    Recognizes high molecular weight and low molecular weight Urokinase.
  • Tested applications
    Suitable for: ELISA, WBmore details
  • Species reactivity
    Reacts with: Mouse
  • Immunogen

    Recombinant full length protein (Mouse).

Properties

Applications

Our Abpromise guarantee covers the use of ab20789 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ELISA
WB
  • Application notes
    ELISA: 1/10000. WB: 1/5000. Predicted molecular weight: 48 kDa. Dilution optimised using Chromogenic detection. Not tested in other applications. Optimal dilutions/concentrations should be determined by the end user.
  • Target

    • Function
      Specifically cleave the zymogen plasminogen to form the active enzyme plasmin.
    • Tissue specificity
      Expressed in the prostate gland and prostate cancers.
    • Sequence similarities
      Belongs to the peptidase S1 family.
      Contains 1 EGF-like domain.
      Contains 1 kringle domain.
      Contains 1 peptidase S1 domain.
    • Post-translational
      modifications
      Phosphorylation of Ser-158 and Ser-323 abolishes proadhesive ability but does not interfere with receptor binding.
    • Cellular localization
      Secreted.
    • Information by UniProt
    • Database links
    • Alternative names
      • ATF antibody
      • ATF uPA antibody
      • BDPLT5 antibody
      • Plasminogen activator antibody
      • Plasminogen activator urinary antibody
      • Plasminogen activator urokinase antibody
      • PLAU antibody
      • QPD antibody
      • u PA antibody
      • U plasminogen activator antibody
      • u-PA antibody
      • U-plasminogen activator antibody
      • uPA antibody
      • URK antibody
      • UROK_HUMAN antibody
      • Urokinase plasminogen activator antibody
      • Urokinase type plasminogen activator antibody
      • Urokinase type plasminogen activator precursor antibody
      • Urokinase-type plasminogen activator chain B antibody
      see all

    References

    This product has been referenced in:
    • Schroder WA  et al. Tumor cell-expressed SerpinB2 is present on microparticles and inhibits metastasis. Cancer Med 3:500-13 (2014). WB . Read more (PubMed: 24644264) »
    • Hasegawa Y  et al. Urokinase-targeted fusion by oncolytic Sendai virus eradicates orthotopic glioblastomas by pronounced synergy with interferon-ß gene. Mol Ther 18:1778-86 (2010). Read more (PubMed: 20606645) »
    See all 2 Publications for this product

    Customer reviews and Q&As

    Question

    BATCH NUMBER 204795 ORDER NUMBER 188849 DESCRIPTION OF THE PROBLEM no band in 48kD in W.B. also can be seen in 30kD, 37kD SAMPLE mouse spleen, kidney, and aorta tissue from C57BL6, uPA-/-, and uPAR-/- mice. PRIMARY ANTIBODY ab20789, 1:5000, diluted in 2% milk TNT buffer incubate for 1 hour in 37C or overnight in 4C DETECTION METHOD Supersignal west PICO Chemiluminescent Substrate, PIERCE POSITIVE AND NEGATIVE CONTROLS USED no-positive control use. Rather, I want to find positive control. ANTIBODY STORAGE CONDITIONS -20C, diplicated 10ul each SAMPLE PREPARATION both reducing and non-reducing condition. reducing buffer is boiled. non-reducing buffer is unboiled. Lysis buffer is from cell-signaling, protease inhibitors cocktail is from Sigma. AMOUNT OF PROTEIN LOADED 10ug or 20ug ELECTROPHORESIS/GEL CONDITIONS I used 10% Gel both reducing and non-reducing condition. 120V in loading for 30-60min TRANSFER AND BLOCKING CONDITIONS 100V in transfer for 90 min. Blocking; 2% milk in TNT buffer for 1 hour in 37C. SECONDARY ANTIBODY HRP goat anti-rabbit IgG, Vector, PI-1000, 1:10,000, diluted in 2% milk TNT buffer,incubate for 1 hour in 37C HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 4 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? No WHAT STEPS HAVE YOU ALTERED? Incubation time; 4C overnight to 37C for 1 hour ADDITIONAL NOTES I have uPA-/- mice and uPAR-/- mice. I removed kidney, spleen and aorta tissue from C57BL6, uPA-/- and uPAR-/- mice. Then I isolated their protein from each samples. Under reducing condition, I could find 30kD bands. But even in uPA-/- sample lane, I could find. Under non-reducing condition, I could find 37kD band in C57BL6 and uPAR-/- mice lane but not in uPA-/- lane.

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    Answer

    Thank you for your enquiry. I am sorry to hear that you have been having difficulties with this antibody. The results that you have been obtaining are a little confusing. You are detecting what appears to be cleavage fragments under reducing and non-reducing condition but strangely you are detecting the cleaved product in uPA-/- tissues under non-reducing conditions. You are preparing and electrophoresing the samples in a manner that I would recommend. We do not sell a positive control lysate or recombinant protein. However, following a brief search I located the following potential positive control of purified recombinant murine Urinary-type plasminogen activator (uPA); http://www.stratech.co.uk/merchant.ihtml?new_id=2101&step=2 This control would give you a good gage to ascertain the migration of high molecular weight uPA. I am in touch with the source of this antibody in order to try to find some additional information with respect to suitable conditions for the application of this antibody. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

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    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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