Overview

  • Product name
    Anti-Thrombospondin 1 antibody
  • Description
    Rabbit polyclonal to Thrombospondin 1
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IHC-P, ICC/IF, IP, WBmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat, Chicken, Cow
  • Immunogen

    Synthetic peptide corresponding to Human Thrombospondin 1 aa 750-850 conjugated to keyhole limpet haemocyanin.
    Database link: 85762
    (Peptide available as ab94880)

  • Positive control
    • This antibody gave a positive signal in HUVEC whole cell lysate. IHC-P: FFPE human breast adenocarcinoma tissue sections. ICC/IF: HeLa cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab85762 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
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF Use a concentration of 5 µg/ml.
IP Use at an assay dependent concentration.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 155 kDa (predicted molecular weight: 129 kDa). Human Thrombospondin-1 is predicted to migrate in WB to ~155kDa as it contains a number of potential glycosylation sites; Swiss Prot predicts a protein of 129kDa not taking modifications into account.

Target

Images

  • Anti-Thrombospondin 1 antibody (ab85762) at 1 µg/ml + HUVEC (Human Umbilical Vein Endothelial Cell) Whole Cell Lysate at 10 µg

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 129 kDa
    Observed band size: 155 kDa
    why is the actual band size different from the predicted?


    Exposure time: 5 minutes


    Human Thrombospondin-1 is predicted to migrate in WB to ~155kDa as it contains a number of potential glycosylation sites; Swiss Prot predicts a protein of 129kDa not taking modifications into account.
  • IHC image of Thrombospondin staining in human breast adenocarcinoma formalin fixed paraffin embedded tissue section, performed on a Leica Bond system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab85762, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • ab85762 staining Thrombospondin 1 in HeLa cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with the antibody ab85762 at 5µg/ml and ab7291 (Mouse monoclonal to alpha Tubulin - Loading Control) used at a 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed, (pseudo-colored green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594) preadsorbed, (colored red), both used at a 1/1000 dilution for 1 hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43 µM for 1hour at room temperature.

  • ab85762 used in Immunoprecipitation.
    Whole cell lysate prepared from human endothelial cells loaded at 600µg total protein.
    Immunoprecipitation step performed using Protein A.
    For IP: ab85762 used at 3.33µg/mg lysate.
    For WB: ab85762 used at a1/1000 dilution.

    See Abreview

References

This product has been referenced in:
  • Smeda M  et al. Dual antiplatelet therapy with clopidogrel and aspirin increases mortality in 4T1 metastatic breast cancer-bearing mice by inducing vascular mimicry in primary tumour. Oncotarget 9:17810-17824 (2018). Read more (PubMed: 29707148) »
  • Anene C  et al. Platelet microparticle delivered microRNA-Let-7a promotes the angiogenic switch. Biochim Biophys Acta 1864:2633-2643 (2018). WB . Read more (PubMed: 29684582) »
See all 20 Publications for this product

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (mouse aorta)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: PH=6 ,citrate buffer
Permeabilization
No
Specification
mouse aorta
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Fixative
10% Formalin

Abcam user community

Verified customer

Submitted Jun 28 2019

Application
Western blot
Sample
Rabbit Cell lysate - whole cell (aqueous humor from rabbit eyes)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Specification
aqueous humor from rabbit eyes
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Xiaoqing Guo

Verified customer

Submitted Apr 30 2019

Application
Immunohistochemistry (Frozen sections)
Sample
Rat Tissue sections (heart)
Permeabilization
Yes - 0.5% triton-x100
Specification
heart
Blocking step
Glycine as blocking agent for 30 minute(s) · Concentration: 3.75µg/mL · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Mar 07 2018

Question
Answer

Thank you for your phone call.

DISCOUNT CODE: xxx
Expiration date: xxx
Value: xxx

m very pleased to hear you would like to accept our offer and test ab85762 in mouse. This code will give you a discount off your next order before the expiration date. To redeem this offer, please submit an Abreview for mouse and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. Please remember that submission of the Abreview is sufficient for the discount code to become active.
For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.

Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

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Read More
Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (endothelial)
Total protein in input
600 µg
Specification
endothelial
Immuno-precipitation step
Protein A

Abcam user community

Verified customer

Submitted Nov 24 2011

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