Anti-Thrombospondin 1抗体(ab85762)

概述

  • 产品名称
    Anti-Thrombospondin 1抗体
  • 描述
    兔多克隆抗体to Thrombospondin 1
  • 经测试应用
    适用于: IHC-P, ICC/IF, IP, WBmore details
  • 种属反应性
    与反应: Mouse, Human
    预测可用于: Rat, Chicken, Cow
  • 免疫原

    Synthetic peptide corresponding to Human Thrombospondin 1 aa 750-850 conjugated to Keyhole Limpet Haemocyanin (KLH).
    (Peptide available as ab94880)

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

性能

应用

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.

应用 Ab评论 说明
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.

靶标

图片

  • 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 minutesHuman 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.

  • ICC/IF image of ab85762 stained HeLa cells. The cells were 4% PFA fixed (10 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 (ab85762, 5µg/ml) 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. This antibody also gave a positive result in 4% PFA fixed (10 min) HepG2 cells at 5µg/ml, and in 100% Methanol fixed (5 min) HeLa, and HepG2 cells at 5µg/ml.
  • 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

文献

This product has been referenced in:
  • Fallahi-Sichani M  et al. Adaptive resistance of melanoma cells to RAF inhibition via reversible induction of a slowly dividing de-differentiated state. Mol Syst Biol 13:905 (2017). Read more (PubMed: 28069687) »
  • Jeanne A  et al. Lumican delays melanoma growth in mice and drives tumor molecular assembly as well as response to matrix-targeted TAX2 therapeutic peptide. Sci Rep 7:7700 (2017). Read more (PubMed: 28794454) »

See all 10 Publications for this product

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (endothelial)
Total protein in input
600 µg
Specification
endothelial
Immuno-precipitation step
Protein A
Username

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提交于 Nov 24 2011

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

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