Anti-Angiopoietin 2/ANG2 antibody (ab8452)

Reviews (4)Q&A (7)References (18)

Overview

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • Storage buffer
    pH: 7.20
    Preservative: 0.1% Sodium azide
    Constituents: 0.42% Potassium phosphate, 0.87% Sodium chloride
  • Purity
    Whole antiserum
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab8452 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/IF 1/200.
IHC-Fr Use at an assay dependent concentration. PubMed: 19553662Use at an assay dependent dilution (PMID 19553662).
WB 1/500. Can be blocked with Mouse Angiopoietin 2/ANG2 peptide (ab9077). There is reaction with serum in the cell supernatants, which results in strong background and makes it difficult to see the angiopoietins in cell supernatants. However when precipitated (using soluble Tie2) the signals are very good and strong.

Target

  • Function
    Can induce tyrosine phosphorylation of TIE2. Binds to TIE2 receptor and counteracts blood vessel maturation/stability mediated by angiopoietin-1. Its function may be context-dependent. In the absence of angiogenic inducers, such as VEGF, ANG2-mediated loosening of cell-matrix contacts may induce endothelial cell apoptosis with consequent vascular regression. In concert with VEGF, it may facilitate endothelial cell migration and proliferation, thus serving as a permissive angiogenic signal.
  • Sequence similarities
    Contains 1 fibrinogen C-terminal domain.
  • Domain
    The Fibrinogen C-terminal domain mediates interaction with the TEK/TIE2 receptor.
  • Cellular localization
    Secreted.
  • Information by UniProt
  • Database links
    see all
  • Alternative names
    • AGPT 2 antibody
    • Agpt2 antibody
    • ANG 2 antibody
    • ANG-2 antibody
    • ANG2 antibody
    • Angiopoietin 2a antibody
    • Angiopoietin 2B antibody
    • Angiopoietin-2 antibody
    • Angiopoietin2 antibody
    • ANGP2_HUMAN antibody
    • ANGPT 2 antibody
    • Angpt2 antibody
    • Tie2 ligand antibody
    see all

Images

  • Western blot - Anti-Angiopoietin 2/ANG2 antibody (ab8452)
    Western blot - Anti-Angiopoietin 2/ANG2 antibody (ab8452)This image is courtesy of Marion Scharpfenecker 2002

    Supernatants of mouse angiopoietin-expressing endothelial cells. Soluble Tie 2 was used to precipitate the angiopoietins to reduce background.

    Lane 1 - mock
    Lane 2 - mouse angiopoietin-2 (clone 2-9) expressing cells
    Lane 3 - mouse angiopoietin-1 (clone 1-15) expressing cells
    Lane 4 - mouse angiopoietin-1 (clone 1-8) expressing cells
    Lane 5 - wt

     

  • Immunocytochemistry/ Immunofluorescence - Anti-Angiopoietin 2/ANG2 antibody (ab8452)
    Immunocytochemistry/ Immunofluorescence - Anti-Angiopoietin 2/ANG2 antibody (ab8452)
    ICC/IF image of ab8452 stained PC12 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab8452, 1/200 dilution) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • Immunohistochemistry (Frozen sections) - Anti-Angiopoietin 2/ANG2 antibody (ab8452)
    Immunohistochemistry (Frozen sections) - Anti-Angiopoietin 2/ANG2 antibody (ab8452)Image from Liu XS et al., J Biol Chem. 2009 Aug 21;284(34):22680-9. Epub 2009 Jun 24. Fig 1.; doi:10.1074/jbc.M109.006551; August 21, 2009, The Journal of Biological Chemistry, 284, 22680-22689.

    Immunohistochemical analysis of murine brain coronal sections, staining Angiopoietin 2/ANG2 with ab8452 at 1/500 dilution. Sections were taken from control mice (left) or mice subjected to middle cerebral artery occlusion (right).

References

This product has been referenced in:
  • Bohn KA  et al. Inhibition of VEGF and Angiopoietin-2 to Reduce Brain Metastases of Breast Cancer Burden. Front Pharmacol 8:193 (2017). IHC-Fr ; Mouse . Read more (PubMed: 28443023) »
  • Rodrigues T  et al. Methylglyoxal-induced glycation changes adipose tissue vascular architecture, flow and expansion, leading to insulin resistance. Sci Rep 7:1698 (2017). WB ; Rat . Read more (PubMed: 28490763) »
See all 18 Publications for this product

Publishing research using ab8452? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-10 of 11 Abreviews or Q&A

Immunocytochemistry/ Immunofluorescence abreview for Anti-Angiopoietin 2 antibody

 Good
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Human Umbilical Vein Endothelial Cell)
Permeabilization
Yes - 0.5% Triton X-100 for 20min
Specification
Human Umbilical Vein Endothelial Cell
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 37°C
Fixative
Formaldehyde
Read More

Abcam user community

Verified customer

Submitted Aug 23 2017

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Angiopoietin 2 antibody

 Poor
Abreviews
Abcam has not validated the combination of species/application used in this Abreview.
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (mouse kidney)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris/EDTA pH 9.0
Permeabilization
Yes - 0.5% Triton X100
Specification
mouse kidney
Blocking step
2.5% BSA / 2.5% donkey serum as blocking agent for 4 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde
Read More

Abcam user community

Verified customer

Submitted Jun 03 2016

Western blot abreview for Anti-Angiopoietin 2 antibody

 Average
Abreviews
abreview image
Application
Western blot
Sample
Mouse Tissue lysate - whole (lung)
Gel Running Conditions
Non-reduced Non-Denaturing (Native)
Loading amount
30 µg
Treatment
OVA I.P, I.N
Specification
lung
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More

Abcam user community

Verified customer

Submitted Oct 05 2015

Western blot abreview for Anti-Angiopoietin 2 antibody

 Below Average
Abreviews
Application
Western blot
Loading amount
50 µg
Gel Running Conditions
Non-reduced Denaturing (10%gel)
Sample
Mouse Tissue lysate - whole (lung)
Specification
lung
Treatment
OVA-induce
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 38°C
Read More

Abcam user community

Verified customer

Submitted Dec 03 2012

Question

Dear technical team,

Our customer directly claimed to US.technical team and received e-mail of below.
So she would like to receive a credit note.
Please check attached Original Message.

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Answer

Thank you very much for your reply.

I will forward your request to my colleagues in our Hong Kong office, and someone will be in touch with you promptly.

Have a nice day!

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Question

AbID: 8452, Anti-Angiopoietin 2 antibody Rating: Below Average Sample: Species: Mouse Type: Tissue lysate - whole Loading amount: 50 µg Specification: lung Treatment: OVA-induce Application: Application: Gel Running Conditions: Non-reduced Denaturing (10%gel) Blocking step: Milk as blocking agent for 2 hours at 38°C Blocking Concentration: 5% Other detail: Dilution: 1/1000 Incubation time: 18 hours at 4°C Diluent: 5% milk Secondary Antibody: Name: Non-Abcam Antibody was used: Goat anti- rabbit lgG-hrp Conjugation: HRP Dilution: 1/15000 Detection: Detection method: ECL Exposure: 1 minute and 0 seconds Bands: Specific: 75 kDa Non-specific: several kDa Additional Data:

Read More
Answer

Thank you very much for your valuable review.
I'm writing to follow-up with you, as the results were below average and we do guarantee this antibody to work in Western blot with mouse samples. I'm sorry to see that there were several non-specific bands on the blot. We have found that ab8452 gives more specific results following immunoprecipitation. I'd be happy to offer you a replacement with a different antibody such as ab125692, or alternatively a credit or refund.
https://www.abcam.com/Angiopoietin-2-antibody-ab125692.html
If you would like to accept one of these options, please let me know your original order or PO number and I'll arrange this for you promptly.
I look forward to hearing from you. If you have any questions or need anything else, please let me know and I'll be happy to help.

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Question

In the western blot shown, you mentioned using sTie2 to precipitate. do you have a protocol for that?

Read More
Answer

Thank you for your enquiry. The Tie2 precipitation experiment was performed by an outside collaborator, and we do not have any further information at hand regarding this. I have tried to email the collaborator to see if we can obtain this information, as several customers have enquired about this. I see that one of my colleagues attempted previously and we did not receive a reply, so I am not optimistic about getting further information. If I do obtain the information you requested, I will forward it along to you.

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Question

How specific are these antibodies to mouse? How specific are they to 1 vs. 2 and vice versa?

Read More
Answer

Thank you for your enquiry. ab8451 is specific for mouse Angiopoietin 1. ab8452 cross-reacts with human (reacts with Ang-2 and weakly with Ang-1) and mouse (specific for mAng-2). There is little sequence homology between the two epitopes of 1 versus 2, therefore, I do not expect either antibody to cross react with the other form. For example, ab8451 is specific for mouse Angiopoietin 1 and should not cross react with mouse Angiopoietin 2. Here is the clustalW alignment of the 2 epitopes: mouseAng2 MWQIIFLTFGWDLVLAS--AYSNFRKSVDSTGRRQYQVQNGP mouseAng1 --MTVFLSFAFFAAILTHIGCSNQRRNPENGGRRYNRIQHGQ :**:*.: .: : . ** *:. :. *** ::*:* I hope this information helps, please do not hesitate to contact us if you need any more advice or information,

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Question

On your website, there is a representative Western blot using the Ang2 ab (ab8452) performed by Marion Scharpfenecker. Do you have the protocol for this procedure? Why is there a band at 50KD? Is the antibody non-specific? Thank you.

Read More
Answer

All the information available for this product is listed on the on-line datasheet (price, datasheet, publication, suitability, cross-reactivity). Angiopoietin-2 is a naturally occurring antagonist of angiopoietin-1. Western blot analysis showed that ANG2 is expressed as a homodimeric 68 kDa protein that is reduced to around 50 kD, after deglycosylation. We would refer you to the extensive “Resources (Popular Protocols)” section on the Abcam website, you may find useful information there.

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Question

Has this antibody been used for IHC on mouse paraffin sections? If so, at what concentration? What type of antigen retrieval is needed, if any? Please forward any pertinant protocol info. thnx.

Read More
Answer

This antibody has never been tested in IHC on mouse tissue sections, therefore we can't guarantee results.

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1-10 of 11 Abreviews or Q&A

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