Overview

  • Product name
  • Description
    Rabbit polyclonal to Angiostatin
  • Host species
    Rabbit
  • Specificity
    Detects recombinant human angiostatin.
  • Tested applications
    Suitable for: ICC, IHC-Fr, WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide corresponding to Mouse Angiostatin aa 98-116.
    Sequence:

    VYLSECKTGIGNGYRGTMS


    (Peptide available as ab4925)

Properties

Applications

Our Abpromise guarantee covers the use of ab2904 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
ICC 1/10 - 1/100.
IHC-Fr Use at an assay dependent concentration. PubMed: 17496220MOPS fixed frozen tissue.
WB Use a concentration of 2 µg/ml. Detects a band of approximately 38, 50 kDa (predicted molecular weight: 38 , 50 kDa).Can be blocked with Plasminogen peptide (ab4925).
ICC/IF Use at an assay dependent concentration.

Target

  • Relevance
    Angiostatin is an internal proteolytic fragment of plasminogen, containing at least three Kringle domains. The Kringle domains have specific anti-migratory and anti-proliferative activities. Angiostatin, an endogenous angiogenesis inhibitor, inhibits endothelial cell proliferation in vitro and angiogenesis in vivo. It is a potent and specific natural inhibitor of angiogenisis and metastatic tumor growth.
  • Cellular localization
    Secreted
  • Database links

Images

  • Immunocytochemistry/Immunofluorescence analysis of Angiostatin (green) showing staining in the cytoplasm of NIH-3T3 cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were incubated with ab2904 in 3% BSA-PBS at a dilution of 1:20 and incubated overnight at 4 ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

  • Immunocytochemistry/Immunofluorescence analysis of Angiostatin (green) showing staining in the cytoplasm of K562 cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with ab2904 in 3% BSA-PBS at a dilution of 1:20 and incubated overnight at 4 ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. Nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

  • All lanes : Anti-Angiostatin antibody (ab2904) at 1 µg/ml

    Lane 1 : Human liver tissue lysate - total protein (ab29889)
    Lane 2 : Human Plasma Total Protein Lysate
    Lane 3 : Human testis tissue lysate - total protein (ab30257)

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 38 , 50 kDa
    Observed band size: 38 + 50 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 27 kDa, 48 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 12 minutes

References

This product has been referenced in:
  • Montagner A  et al. Src is activated by the nuclear receptor peroxisome proliferator-activated receptor ß/d in ultraviolet radiation-induced skin cancer. EMBO Mol Med 6:80-98 (2014). IF ; Mouse . Read more (PubMed: 24203162) »
  • Wu T  et al. Urinary angiostatin--a novel putative marker of renal pathology chronicity in lupus nephritis. Mol Cell Proteomics 12:1170-9 (2013). Human . Read more (PubMed: 23345539) »
See all 9 Publications for this product

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

Application
Western blot
Sample
Mouse Tissue lysate - whole (heart)
Gel Running Conditions
Reduced Denaturing (10)
Loading amount
10 µg
Specification
heart
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Abcam user community

Verified customer

Submitted Sep 26 2017

Application
Western blot
Sample
Mouse Tissue lysate - whole (lung)
Gel Running Conditions
Non-reduced Non-Denaturing (Native)
Loading amount
30 µg
Specification
lung
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Oct 05 2015

Answer

The antibody recognizes angiostatin: plasminogen (as well as PLC and Microplasmin) are synonyms for angiostatin.

I hope this answers your question. Please feel free to contact us with any additional questions.

Read More

Answer

From the reading of references which have used this antibody I would expect this antibody to have been used in a reduced form as the materials and methods for the following reference state they performed a standard WB (Balaggan KS et al. EIAV vector-mediated delivery of endostatin or angiostatin inhibits angiogenesis and vascular hyperpermeability in experimental CNV. Gene Ther 13:1153-65 (2006). PubMed: 16572190). We also recommend to use 5% Non-fat dry milk to block the membrane and to incubate the primary antibody in TBST with milk as well. Please do not hesitate to contact us if your customer has further questions.

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Answer

Thank you for your enquiry. Regarding the polyclonal antibody to Angiostatin (ab2904), this has not been tested for cross-reactivity with plasminogen at this time. I would assume if the antibody detects angiostatin, it should also bind to plasminogen since angiostatin is a cleavage product of plasminogen. At this time, however, the antibody has only been tested on recombinant samples. I am looking into having this datasheet changed to reflect that the antibody was targeted specifically to angiostatin and not plasminogen. I hope this information helps. Please do not hesitate to contact us again if you need anything further.

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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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