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

Anti-Thrombospondin 1 antibody 图像

  • 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.

  • 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.
  • 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

Anti-Thrombospondin 1 antibody (ab85762)参考文献

This product has been referenced in:
  • Caolo V  et al. ADAM10 and ADAM17 have opposite roles during sprouting angiogenesis. Angiogenesis 18:13-22 (2015). Mouse . Read more (PubMed: 25218057) »
  • Pan XY  et al. Hirudin promotes angiogenesis by modulating the cross-talk between p38 MAPK and ERK in rat ischemic skin flap tissue. Tissue Cell 47:301-10 (2015). Rat . Read more (PubMed: 25958163) »

See all 7 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
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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"